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An Introduction to


Pioneering science delivers vital medicines

Amgen is a leading human therapeutics company in the biotechnology industry. Since our founding in 1980, we have focused on accomplishing our mission to serve patients by discovering, developing and delivering innovative medicines to treat grievous illnesses. By pioneering the development of novel products based on advances in cellular and molecular biology, Amgen’s therapeutics have changed the practice of medicine and helped millions of people around the world to fight cancer, kidney disease, rheumatoid arthritis and other serious illnesses.

Table of Contents
Chapter One: Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Chapter Two: The Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Chapter Three: How Biology Drives Biotechnology . . . . . . . . . . 9 Chapter Four: The Technology . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Chapter Five: Drug Discovery . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 Chapter Six: Drug Development . . . . . . . . . . . . . . . . . . . . . . . . . 22 Chapter Seven: Scale-Up and Manufacturing . . . . . . . . . . . . . . 26 Chapter Eight: Biotechnology Medicines . . . . . . . . . . . . . . . . . . 29 Chapter Nine: Future of Biotechnology in Healthcare . . . . . . . . 31 Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Timeline of Medical Biotechnology . . . . . . . . . . . . . . . . . . . . . . 42

Chapter One:


In 1919, Karl Ereky, a Hungarian engineer, coined the term biotechnology* to describe the interaction of biology and human technology. He envisioned a new era of technology based on using biology to turn raw materials into socially useful products. Nearly a century later, Ereky’s vision is being realized by thousands of companies and research institutions.



*Terms in boldface are defined in the glossary .

Modern biotechnology began in the 1970s in Northern California and has since grown into a worldwide industry . Amgen, founded in 1980, was one of the first companies to realize the new field’s promise by bringing biotechnology medicines to patients . Today, biotechnology industry sectors include healthcare (biologics, devices, diagnostics), agriculture (genetically modified organisms, food safety), industry and environment (biofuels, biomaterials, pollution) and biodefense (vaccines, biosensors). This booklet focuses on healthcare . Drugs A drug is a therapeutic substance used to prevent, manage or cure disease . In the United States, the U .S . Food and Drug Administration (FDA) must approve all drugs before they are sold to the public . Most countries follow global harmonized guidelines and have a regulatory agency similar to the FDA that evaluates drug research and approves drugs for marketing . The most familiar type of drug is the synthesized drug, such as aspirin . The pharmaceutical industry traditionally manufactures synthesized drugs . The biotechnology revolution brought about a new class of drug: the biologic . Biologics are therapies derived from living organisms and include therapeutic proteins, DNA vaccines, monoclonal antibodies and peptibodies [a modality that combines the active portion of a protein (peptide) with a portion of the core structure of an antibody], as well as new experimental modalities such as gene therapy, stem cell therapy, antisense nucleotides and RNA viruses .

Many biotechnology drugs are proteins. Proteins, which are made from amino acids, are the workhorses of the cell and perform all functions within a cell . Because cells produce proteins naturally, the biotechnology industry utilizes cells, not chemicals, to manufacture biologics .
BIoFaCT To bring a new drug to market (from discovery through clinical trials and FDA approval) costs an estimated $1 billion and can take 10 to 15 years or longer .* Only one in 10 new drugs that makes it into human testing actually makes it to market . Given this high failure rate and the tremendous cost of bringing a new therapy to market, companies depend on successful drugs to produce enough revenue to compensate for both the R&D costs of the successful therapies and the expense of failed ones .

* Innovation .org . (February 2007) . Drug discovery and development: Understanding the R&D process [Brochure] . Washington, DC: Pharmaceutical Research and Manufacturers of America .



such as humans. The cell is the basic unit of life. such as bacteria and yeast. All living organisms consist of one or more cells. Some organisms are unicellular. All cells have common processes they perform in order to survive. Others. consisting of trillions of cells. Biotechnology harnesses these processes to make products to treat illness and improve health.Chapter Two: The Science The biotechnology industry is based on living organisms. 4 The Science . are multicellular.

If a receptor is blocked. the cytoplasm (which includes the organelles) and the nucleus. and each performs a specific function . It is surrounded by a membrane that lets certain molecules into and out of the nucleus to Ribosomes can assemble an average-size protein in one minute . For example. solute concentration. When a specific type of molecule attaches to a receptor. creating a cellular response . The cell divides and forms two identical cells from the original single cell . Vacuoles store cellular waste products for disposal . Receptors cross the membranes and act as docking stations for molecules . a cell makes a copy of its DNA and other cell parts . The endoplasmic reticulum folds and transports certain proteins . This is how some biologic therapies work: they attach to receptors and interrupt cell signaling . Embedded in the cell membrane are receptors. Cells Grow: After replicating. oxygen supply and the presence of hazardous agents . ribosomes make proteins . For example. a series of chemical reactions (cell signaling) can occur in the cell. BIoFaCT the nucleus . This DNA never leaves Cells Metabolize: Cell metabolism is the process by which cells process nutrients and maintain a living state . Golgi bodies modify proteins and are involved in their transportation around the cell . Cells Adapt: Organisms may thrive or die based on their ability to adapt to adverse environmental conditions such as changes in temperature. the daughter cells grow to their intended size . plants grow toward a light source because light is needed for photosynthesis and the production of energy . Other biologics work by mimicking the signaling molecule . The Science 5 . Light and the ability to respond to its presence are essential to a plant’s survival . Cells Respond to Stimuli: Unicellular and multicellular organisms respond to internal and external stimuli . Cell Membrane Nucleus Nucleus Cytoplasm Cytoplasm Cell Membrane keep the DNA safe . Parts of an Animal Cell The cell can be divided into three main sections: the cell membrane. Cells can respond to a whole range of stimuli . cell signaling is stopped and no response occurs . It monitors what goes into and out of the cell . Mitochondria make energy .Cell Processes Cells Replicate (mitosis): Prior to dividing. Nucleus: The nucleus houses most of the DNA and is the control center of the cell . These identical cells are referred to as daughter cells . Cells break down large molecules into smaller molecules to produce energy and molecular building blocks. Organelles: There are many different types of organelles within the cytoplasm of a cell. which are used to create new cell structures and control cell function . Cell Membrane: The cell membrane is the border that surrounds a cell .

T A T A Chromosome Gene C T A A A T T C G G C A T C A A T T G A T A T C C G G C G C G G DNA The molecular structure of DNA—the double helix. 6 The Science . The order of the As. a phosphate group and a base . eye and muscle movement functions . Mutations in mitochondrial DNA can lead to illnesses such as Kearns-Sayre syndrome. DNA is arranged in large segments called chromosomes. Gs and Most cells within an organism contain the exact same DNA. BIoFaCT Mitochondria are the only organelles in animal cells to have their own DNA . where there may be more or less than the normal 46 chromosomes. The diversity of organisms is a result of the limitless combinations of bases—As. Within a chromosome are specific pieces of DNA called genes. Each nucleotide consists of three components: a deoxyribose sugar. deletions or even inversions of sections of certain chromosomes . a DNA segment would look something like a ladder with two side rails (phosphate and ribose groups) and rungs (base pairs) between them . and the order of the nucleotides determines the information stored . Genes vary in length and contain the information a cell needs to make proteins . In each human cell. Ts. Wilkins’s colleague Rosalind Franklin. The As bond with Ts. guanine (G) and cytosine (C) . Proteins control many aspects of cellular function . or structural abnormalities. who died from cancer at the age of 37. whose X-ray crystallography images of DNA helped them clarify the structure of DNA. where there may be duplications. Ts. for a total of 46—half inherited from the mother and half from the father . which causes the loss of heart. Francis Crick and Maurice Wilkins jointly received the Nobel Prize in Physiology or Medicine for discovering the structure of deoxyribonucleic acid (DNA). translocations. DNA is very long. Because the Nobel Prize is awarded only to the living. But many attribute the success of Watson and Crick’s 1953 discovery to Franklin. The resulting A-T and C-G combinations are called base pairs. There are four different types of bases: adenine (A). The structure makes DNA very stable and able to carry vast amounts of information . the Humans have 23 pairs of chromosomes. but not all genes within each cell are active. James Watson. could not be honored. to a story in which the word appears . the length of DNA is equal to 3 billion base pairs . the information encoded by the gene is used to produce. BIoFaCT The information in DNA is stored as a code made up of four basic “building blocks” called nucleotides . and the Cs bond with Gs . When a gene gets turned on. Every organism contains DNA. If flattened. DNA is called a double helix because it is posited to consist of two strands of nucleotides that bond together in a very specific manner . by extension. DNA is the blueprint for the construction and operation of the cell . thymine (T).Determining the Structure of DNA In 1962. Chromosomal abnormalities can involve numeric abnormalities. but the number and arrangement of bases are different for every organism . or turned on . DNA All cells contain deoxyribonucleic acid (DNA). Cs . or express. Gs and Cs in DNA gives meaning to the cell. just as the order of letters in a word gives meaning to that word and.

National Center for Biotechnology Information Established in 1988.S. Mutations also can occur during the natural process of DNA replication. For example. occasionally. Many diseases are the results of genes improperly turned on or off . the National Center for Biotechnology Information (NCBI) creates and maintains extensive networks of biomedical databases for storing and analyzing genomes. The order of amino acids is determined by the DNA sequence in a gene . the number of base pairs and genes does not correspond to the intelligence or physical capability of an organism . National Library of Medicine at the National Institutes of Health. genes and proteins. the rate is one in every 1. Most of these mutations. When a gene is not active. the two are indepenUntwist A T C G G C T A A T C G G C T A A T C G G C T A A T C G G C T A A T C G G C T A A T G C The U. has the largest-known genome—which may be as large as 670 billion base pairs—as compared to the human genome of 3 billion base pairs . See http://www. intelligent and physically capable organism . do not adversely affect us. of the protein . Furthermore.200 base pairs . The human genome is the entire DNA content found in a human. 97 percent is termed noncoding DNA—in other words.ncbi. Genomes The term genome refers to the entire genetic information in an organism .The number of base pairs does not correspond to the number of genes. Biologists have not yet fully discovered the function of noncoding DNA. The databases include information about various genome base pair sequences. Ts. and so on .300 base pairs in most organisms . it is not used to express proteins . Mutations Any change in the DNA sequence is called a mutation . The differences between genomes lie in the number and sequence of base pairs and the number and sequence of genes . Depending on the cell’s function and needs.nih.000 to 25. Gs and Cs . All genomes are made up of the same bases: As. BIoFaCT Read GATATTTCAGGCGCGTCAA CTATAAAGTCCGCGCAGTT encoded protein .gov. Genetic diversity results from an accumulation of mutations over a long period of time. deleted or repeated .000 genes . but they suspect it may be involved in the evolution of species or may have regulatory functions within the cell . Changes in the DNA sequence may cause proteins to become dysfunctional or may even. Environmental factors such as exposure to radiation or chemical toxins can cause mutations . or is turned off. genes are either turned on or off .nlm. An amoeba. and therefore the function. Only 3 percent of the human genome codes for genes. Proteins Proteins are made of long chains of amino acids that fold into intricate and complex 3-D shapes . the corn genome is the entire DNA content found in corn. a unicellular organism. There are 20 different amino acids . dent of each other . The Science 7 . NCBI is a division of the U. the human genome has 3 billion base pairs and approximately 20. improve function .S. Yet a human is a more complex. which causes the differences among species . Compare the human genome with the amoeba genome . however. DNA that does not contain instructions for creating proteins . when the cell is responsible for copying 3 billion base pairs in 20 hours . Bases can be substituted. The sequence of amino acids determines the shape. Variations in individual nucleotides occur within DNA at the rate of approximately one in every 1. In humans. only a few are involved in the production of dysfunctional proteins or disease states .

are single-stranded and use U. Ribosomes will bind to mRNA . Transcription During transcription.BIoFaCT The highest-known number of genes in an organism is around 60. the amino acids are linked together in a very specific combination as dictated by the sequence of nucleotides on the mRNA . Short chains of amino acids are called peptides. Proteins called signaling molecules allow cells to relay messages to one another . 8 The Science . or thymine. Protein receptors receive signals through a type of communication known as signal transduction or cell signaling . Proteins are involved in cellular recognition and identify different types of cells . Proteins called enzymes put molecules together and/or break molecules apart .000—for the bacterium that causes trichomoniasis— which is almost three times as many as in the human genome . Some proteins. Structural proteins give shape to cells and organisms . During translation. Polypeptides fold into a functional protein . transfer RNA (tRNA) reads mRNA and picks up the corresponding amino acids . the original DNA code is rewritten onto a molecule called messenger RNA (mRNA). as one of the four possible bases . Hundreds of different types of proteins perform specific jobs within and between cells . multistep process . are involved with defending an organism against disease . the ribosomes assemble individual amino acids into proteins . instead of T. such as antibodies. Long chains of amino acids are called polypeptides . Every three mRNA nucleotides make up a codon—the code for an amino acid . In this way. Translation During translation. Two of the steps involved with making a protein are transcription and translation. Protein synthesis is a complicated. Some proteins move substances into and out of the cell . RNA molecules are very similar to DNA but have a ribose sugar. or uracil. mRNA is also made up of a sequence of nucleotides that differ slightly from DNA nucleotides .

Chapter Three: How Biology Drives Biotechnology Biotechnology is based on biology. proteins and cell parts to pinpoint the differences between diseased and healthy cells. The basic unit of life is the cell. Biologists study the structure and functions of cells—what cells do and how they do it. they are better able to develop innovative medical diagnostics. which is the study of life. How Biology Drives Biotechnology 9 . Biomedical researchers use their understanding of genes. devices and therapies. Biotechnologists use this information to develop products. When researchers know how diseased cells are altered and when they learn how to affect those alterations.

One approach is to obtain samples of diseased cells and healthy cells and grow them using a method called cell culture. what genes are turned on or turned off in the diseased cells? What proteins are produced—or not produced—in diseased cells as compared to healthy cells? If the disease is caused by a pathogen. psoriatic arthritis. By studying how cellular processes differ between healthy and diseased cells. Models for Studying Disease Researchers often take several different approaches to creating models for studying a particular disease . Take treatments available for autoimmune disorders. fever. To design and develop new drugs. degenerative inflammatory arthritis primarily affecting the spine and causing eventual fusion of the vertebrae. what is the interaction between the pathogen and the person? BIoFaCT The biotechnology industry is one of the most R&D-intensive industries in the world . psoriasis. anemia and hair loss. When too much TNF is produced. for example . An understanding of fundamental biology may lead to effective therapies for patients . The United States is recognized as leading the world in biotechnology R&D . Autoimmune disorders occur when a person’s immune system overreacts and attacks proteins. researchers hope to come to understand the mechanism of disease . Biotechnology companies have worked to develop medicines that inhibit the activity of TNF . Psoriasis is a noncontagious disorder that causes red scaly patches to appear on the skin. painful. In culture. applied mathematics and statistics . Another approach involves studying shared or similar genes and protein equivalents in other species . joints. Biotechnology medicines are often created specifically to address a particular disease mechanism . Some initial questions researchers Autoimmune Diseases Rheumatoid arthritis is a chronic autoimmune disease that causes inflammation and tissue damage in joints and tendons. ask to understand the underlying mechanisms of a disease are: How does a person get the disease? Which cells become diseased? Is this disease caused by genetics. BIoFaCT The process of analyzing and interpreting biologic scientific data is called computational biology and involves computer science. cells and tissues in the body. Studying disease mechanisms provides researchers with information that can lead them to identify targets for the early stages of the drug discovery process . It can be a disabling and painful condition and can lead to substantial loss of functionality and mobility. often leading to inflammation . Its symptoms can include rash. genes and proteins found in humans are also found in other organisms . excessive inflammation occurs—and that can be damaging to joints. Biologists have learned that tumor necrosis The functions of human genes have been revealed by studying parallel genes in 10 How Biology Drives Biotechnology . Psoriatic arthritis is a type of inflammatory arthritis that affects up to 30 percent of people suffering from psoriasis. These patches are areas of inflammation and excessive skin production. juvenile idiopathic arthritis and ankylosing spondylitis . researchers must understand the disease mechanisms involved . aches. Lupus can affect the kidneys. blood and skin. the cells do what cells do—divide and express genes to produce proteins . Since all organisms are made of cells and all cells perform many similar functions. Juvenile idiopathic arthritis is the most common form of chronic inflammatory arthritis in children. factor (TNF) plays a major role in regulating inflammation .Understanding Disease Mechanisms Early-stage drug research and development (R&D) may begin with understanding the underlying biology of a particular disease . This calls for cells to be incubated and fed with specialized growth media. among other organs. Researchers know that too much TNF is produced in autoimmune disorders such as rheumatoid arthritis. Ankylosing spondylitis is a chronic. Lupus is a chronic inflammatory disease that most commonly affects women of childbearing age. and if so. skin and other parts of the body .

These tests look for DNA fragments identified as causing disease or associated with susceptibility to a disease . BIoFaCT Genetic biomarkers are used in developing diagnostic tests for detecting certain genetic diseases . bioinformatics has evolved to focus on nucleotide sequences. the laboratory mouse and many other organisms for use in comparative genomic studies with the human genome . The scientific community generates volumes of biological data daily . Biomarkers also can be used to predict prognosis—for example. Researchers have obtained complete genomic sequences for the bacterium Escherichia coli (E. This leads to the development of diagnostic tools. The challenge for computer programmers is to design databases that allow researchers to easily access existing data as well as submit new data . By testing for these mutations. Today. Because of technological advances in biotechnology. genes and amino acid sequences . organize and index scientific information so researchers can better understand biology . therapies and preventive medicines . Amgen scientists were the first to show in a phase 3 clinical trial that patients with mCRC carrying specific cancerpromoting mutations in a gene known as KRAS are unlikely to benefit from a drug designed to bind to epidermal growth factor receptor (EGFR). KRAS research is regarded internationally as an important step toward personalized medicine for cancer treatment. coli). This approach has added to our understanding of how specific genes and proteins direct the functioning of human cells—both healthy and diseased . Biomarkers Biomarkers are substances that can be measured and evaluated to indicate (or serve as markers of) normal biologic processes.nonhumans . disease processes or biologic responses to therapeutic treatment and disease intervention . the fruit fly Drosophila melanogaster. a protein that if elevated. the roundworm Caenorhabditis elegans. computer science and information technology into one discipline . Historically. Bioinformatics Bioinformatics combines biology. physicians can now eliminate anti-EGFR antibodies as a treatment option for patients with mutated KRAS tumors. KRAS: A Biomarker Breakthrough A biomarker can now help physicians determine which patients will not respond to a certain class of anticancer agents used to treat metastatic colorectal cancer (mCRC). how a disease is likely to progress if left untreated . Biotechnology companies use this information to form a comprehensive picture of normal cell activity so researchers can better study diseased cells . Once a biomarker is validated (meaning all or most patients with the disease test positive for the specific biomarker). Identifying biomarkers that indicate specific disease is a crucial step in the R&D process for developing new biotechnology diagnostic tools . may indicate prostate cancer . disease may also be detected using molecular biomarkers such as prostatespecific antigen. it can be used to diagnose disease risk or presence of disease or to help doctors determine patient treatment . How Biology Drives Biotechnology 11 . biomarkers were physiological indicators—such as blood pressure or heart rate . BIoFaCT Comparative genomics is the study of genome structure and function among different species . the yeast Saccharomyces cerevisiae. The goal of bioinformatics is to capture.

Proteomics The entire set of proteins produced by an organism is called its proteome . Leukemia is cancer that starts in blood-forming tissue—such as bone marrow—and that causes large numbers of abnormal blood cells to be produced and to enter the blood. brain. diabetes. sarcoma. breast. HIV/AIDS and autoimmune disorders .) that affect many types of tissue (prostate. As the cancer cells replicate and grow. fat. a mass (tumor) is formed . Lymphoma is a cancer of the lymphatic system—a network of thin vessels and nodes throughout the body that helps to fight infection. hormonal change and stress . etc. A drug target is the molecule the drug interacts with to bring about the desired change . The biotechnology industry has contributed significant advances in cancer treatment by developing hormone therapies. a type of white blood cell found in the immune system. melanoma. This is very important because response to a drug can vary between patients: some require higher or lower doses for the the drug to be effective . etc. Cancer biology is a specific field that explores all aspects of the cancer cell . uncontrolled and rapid cell growth. Proteins produced in any specific cell of an organism can vary with a number of factors such as time. lymphoma. Biomarkers are also used in drug discovery to determine whether a drug is effective in animal models and at what doses effectiveness is reached . Lymphoma involves lymphocytes. Sarcoma is cancer that begins in bone. biologics and targeted therapies such as monoclonal antibodies . Proteomics researchers identify all proteins involved with protein synthesis and protein folding . Proteins are common drug targets because they are responsible for most normal cell functions—and malfunctions. Cells from the tumor can break away (metastasize) and spread through the bloodstream or lymphatic system to other parts of the body. ovaries.Researchers study modifications and changes that occur during cellular processes—such as protein synthesis—in various disease states . Cancer starts with changes in one cell or a small group of cells that reproduce uncontrollably . Cancer: From Biology to Treatment Cancer research is a field that has been at the forefront of utilizing bioinformatics. biomarkers can be used in disease management to determine whether a drug is having the desired effect and whether the correct dose is being used . cell division and growth are tightly regulated . their 3-D shape in order to correctly function . At least 80 percent of all cancers are carcinomas. function and interaction within and between cells is crucial for drug discovery . creating new tumors . The majority of these products are therapeutic proteins . muscle. leukemia. Carcinoma is cancer that begins in the skin or in tissues that line or cover internal organs. cartilage. Traditionally.). This is sometimes called hitting the target . skin. Melanoma is a cancerous (malignant) tumor that begins in the cells that produce skin coloring (melanocytes). lungs. Proteins need to fold correctly into BIoFaCT Biotechnology has created more than 200 new biotherapeutics and vaccines. Even small structural defects during the folding process can lead to a number of protein diseases . Understanding protein structure. as in the case of disease . blood vessels or other connective and supportive tissue. including products to treat cancer. In healthy cells. Melanoma is almost always curable in its early stages but can be lethal in later stages. 12 How Biology Drives Biotechnology . radiation and chemotherapy . cancer cells keep dividing—without normal checks and balances—creating greater opportunity for mutations . However. Proteomics is the study of protein structure and function . Proteins control all aspects of cellular function . Types of Cancer Cancer is a general term used to describe diseases that occur due to abnormal. biomarkers and protein studies to develop new therapies that target specific cellular processes . This can lead to the identification of biomarkers that signal a change in disease progression and can be used as a key development in drug discovery and testing . A mutation may result from the environment or could have occurred during DNA replication . There are many types of cancer (carcinoma. Finally. cancer has been treated with surgery. Researchers follow cancer pathways and determine the molecular basis of cancer as they develop diagnostics and treatments .

This section focuses on some of these platform technologies. it’s helpful to have some basic knowledge of what goes on in the lab. To understand the industry.Chapter Four: The Technology Biotechnology scientists depend on a wide variety of constantly evolving laboratory techniques and tools. The Technology 13 .

which is used to physically carry DNA into a host cell . a restriction enzyme found in Haemophilus-aegyptius. The process is called transformation. For example. nonfunctional pieces. All restriction enzymes are specific and reproducible. recognizes and cuts at the six-base sequence GAATTC . recognizes and cuts at the four base sequence GGCC .Transformed Cells Transformation of bacterial cells for the production of recombinant proteins usually involves E. BIoFaCT BIoFaCT Scientists have identified more than 3. called transfection. Restriction Enzymes Biotechnology employs a process called genetic engineering. The ability to cut and paste DNA is the basis of genetic engineering . Cells that contain recombinant DNA are referred to as genetically modified. All DNA. thereby protecting the bacterium from an invading virus . A phage is a genetically engineered virus that injects DNA into bacteria . Daniel Nathans. Transformation of animal cells. insect or mammalian . A host cell can be bacterial. Common bacterial vectors include plasmids and phages . HaeIII. + Restriction enzymes are proteins that function as molecular scissors and cut DNA. the new DNA is called recombinant DNA. Recombinant DNA When segments of DNA are cut and pasted together. Recombinant DNA can be inserted into cells to produce cells with new characteristics .800 restriction enzymes. 14 The Technology . The process utilizes restriction enzymes . A plasmid is a circular unit of DNA that can be engineered to carry a gene of interest . coli. Scientists discovered restriction enzymes (endonucleases) in bacteria in the 1970s . They found that these enzymes cut up viral DNA into small. yeast. EcoR1. a restriction enzyme found in E. Ts. HaeIII reads any DNA segment and cuts the DNA every time it encounters the sequence GGCC . which combines DNA sequences in order to produce recombinant proteins as potential therapeutics . There are hundreds of restriction enzymes— each of them recognizing a specific sequence of DNA called a restriction site . DNA ligase is a protein (enzyme) that seals two DNA segments together in a process called ligation . Gs and Cs . regardless of where it comes from. Recombinant DNA can be introduced into a host cell by a vector. Werner Arber and Hamilton Smith received the 1978 Nobel Prize in Physiology or Medicine for their discovery of restriction endonucleases . coli. is made up of the same four bases—As. plant. The counterpart to cutting is called pasting . Their discovery led to the development of recombinant DNA technology . CHO cells were introduced in the 1960s and remain the most commonly used mammalian host cells for industrial production of recombinant protein therapeutics. usually involves a cell line derived from Chinese hamster ovary (CHO) cells. and more than 600 are commercially available for purchase from scientific supply companies . which are two key characteristics that allow researchers to utilize restriction enzymes to manipulate DNA . This genetic altering can include a single-base (letter) change or multiple gene changes . transgenic or transformed cells .

purified and analyzed for activity and quality before it goes to market . Many human proteins are glycosylated. monoclonal antibodies and hematopoietic growth factors for the treatment of cancer. glycosylated are made in animal cell cultures . Culture media provides all the nutrients necessary for cell growth . Typically. AIDS. Growing large quantities of transformed cells is a major step in the process of producing recombinant protein products . Adding a phosphate group—a process known as phosphorylation—can act as an on-switch. Since only a limited number of cells can be grown in small petri dishes or flasks. because variation in these conditions may affect the proteins expressed and. hormones. Cell Culture Cell culture is the technique of growing cells in the laboratory under controlled conditions . it may not function properly . When recombinant proteins are produced for use as human therapeutics. If a protein is translated but not correctly glycosylated. therefore. meaning. the cell culture can be transferred to large containers called bioreactors. Sometimes further processing is required to yield an active or fully functioning protein . allowing the proteins to become active proteins . asthma and many other conditions . they have a particular pattern of sugar molecules linked to them . Recombinant proteins for therapeutic use include vaccines. It is very important to maintain specific conditions for cultures. often requiring certain gas mixtures of oxygen and carbon dioxide . more-complex proteins that are. Simple proteins can be produced using recombinant DNA technology in bacterial cell cultures . In the process of commercial production of proteins. aller- The Technology 15 . Producing a protein with the proper order of amino acids isn’t always the whole story . the product that is produced . Recombinant Proteins Recombinant DNA can be used to produce recombinant proteins. Other biochemical functional groups may be added to the protein to allow it a larger range of function .BIoFaCT The FDA approved human insulin in 1982— the first medicine made via recombinant DNA technology . Both transformed bacterial cells and transformed animal cells are used in this process . The cultures are grown in an incubator that maintains the appropriate temperature and environment. cell cultures must be scaled up to produce enough protein to meet demand . which are involved in the manufacturing process . The number of recombinant proteins has increased greatly in recent years as the technology used for their production and purification has advanced . During cell culture. The recombinant protein is isolated. cells are grown in petri dishes or flasks containing liquid media . gies. host cells must be grown in large quantities so that enough recombinant protein is produced to meet demand . The host cells use the new DNA information and their own cellular machinery to produce the protein encoded by the recombinant DNA . for example.

Each is based on the type of separating material used. DNA Microarrays A DNA microarray (also called a gene chip) is a small piece of glass or silicon divided into thousands of sections in a grid pattern . Here are a few pieces used in genetic engineering: Thermocycler Polymerase chain reaction (PCR) is a technique that replicates DNA in a machine called a thermocycler. BIoFaCT Kary Banks Mullis developed the polymerase chain reaction in 1983 and received the 1993 Nobel Prize in chemistry for the invention . PCR is a series of cycles that takes a small amount of original DNA and exponentially copies. Gel electrophoresis allows DNA fragments to be separated within a gel . causing it to migrate toward the positive pole of the gel apparatus . it . The individual’s DNA must match the attached gene fragment exactly to bind. Each three-step cycle doubles the amount of DNA present . There are different types of electrophoresis . With the aid of a computer. or amplifies. A single piece of DNA can be turned into millions of copies of the same DNA piece . the double-stranded. Gel Electrophoresis Gel electrophoresis is a technique commonly used in the laboratory for analyzing DNA fragments . Gel electrophoresis has many applications in both clinical and research labs . The tagged DNA is washed over the microarray . Larger fragments of DNA move more slowly than smaller fragments because they encounter resistance from the gel matrix . gels being one type. There are many applications for PCR. A gel electrophoresis apparatus holds the gel and allows electricity to run through it . 16 The Technology . Each DNA fragment is negatively charged. DNA. to become doublestranded DNA . PCR enables researchers to make enough DNA to work with in the lab . checking to see whether the reaction generated the correct DNA fragment . The individual’s DNA binds to any complementary DNA sequences on the slide. DNA from an individual is separated into single strands and tagged with a fluorescent dye . and the types of molecules being separated . including producing enough of a DNA sequence or gene for use in creating recombinant proteins . Each section has a single-stranded gene fragment corresponding to either a healthy or diseased gene . One common use is for verifying PCR products—that is. RNA and proteins can all be separated using electrophoresis in an appropriate apparatus . if they are present. thereby indicating the presence of healthy or diseased DNA . fluorescently tagged DNA spots can be located and measured . It’s like a photocopier for DNA .Research Tools Biotechnology professionals depend on leading-edge laboratory equipment . - + Well Gel Gel electrophoresis.

There are both public and private microarray databases . Microarrays are used for genetic testing. and in the discovery of potential drug targets . Other microarrays include protein. The Technology 17 .Microarray. Microarrays are powerful tools allowing researchers to analyze thousands of genes in one test . tissue. for comparison of genetic information from different individuals or species. analysis and interpretation . chemical compound and antibody microarrays—all of which allow researchers to analyze thousands of data points at one time . BIoFaCT Microarrays are capable of generating so much data that special microarray databases are needed for storage. searches. Microarrays can also be used for researching and identifying genes of interest to be used with recombinant DNA technologies .

they specifically look for ways to change one or more molecular or cellular processes that occur in the affected cells of a diseased tissue or organ.Chapter Five: Drug Discovery During drug discovery. 18 Drug Discovery . scientists search for molecules—either chemical or biological agents—that could alter a disease pathway. As part of the discovery process.

Moreover. Scientists estimate that about 8. if any? Does the company have the expertise. cell surface receptors or signaling pathways within a cell . mass spectrometry (MS). There are various causes of diseases . Target Discovery After identifying an unmet medical need and deciding whether it fits within the company’s portfolio. scientists will Researching the genetic and molecular basis of a disease is called studying the mechanism of disease . a difference in the expression or in the sequence of genes results in abnormal functioning of a person’s cells .000 known therapeutic targets exist today . the pathogen produces molecules that can damage the host organism’s cells . While this sounds straightforward. These libraries are often proprietary and constructed by a company explicitly to support its drug discovery programs. other times it could be deficient or missing . or a method able to detect the difference may not yet have been invented . are also taken into consideration . “What are the differences between healthy and diseased cells?” Ultimately. the top five diseases causing death in the United States in 2005 were heart disease. What is known about the disease? What are the current treatment options. The sources of the compounds found in the drug candidates are often natural products (from microbes. DNA sequencing and computerized imaging . Combinatorial chemistry increases the potential of chemical libraries by synthesizing larger. Drug Discovery Drug Discovery 19 . such as regulatory constraints. scientists look very closely at the biology behind the disease . even millions. scientists have to carefully choose the target . chemists developed huge libraries of chemical compounds—thousands. ask. the pathogen will. of chemicals with different structures used to screen for new drugs. the difference between healthy and diseased cells can be too minute to easily detect. For a disease caused by an external pathogen. Targets can be secreted factors. To select a target. It’s also very important to ensure that the potential benefits of a drug are appropriately weighed against any risks such as possible side effects . display molecules in the infected individual that are not present in a healthy person . In inherited diseases. The goal in target discovery is to identify those different molecules . itself. Moreover. BIoFaCT Chemical Libraries In the 1990s. Initial screening of drug candidates is relatively simple. Combinatorial chemistry for small-molecule drugs includes the synthesis of large organic molecules by adding together smaller organic molecules. Once potential drug candidates or leads are identified. So the scientists will need to decide if the goal will be to block the target or to enhance or replace it in order to restore healthy function . This can be done using a variety of technologies such as microarray experiments. There may be one or more mechanisms of the disease and many points in the mechanism at which to intercede . stroke. Where can they intervene. plants and simple marine life) or chemical compounds synthesized by an organic chemist. Sometimes this leads to a target being present in excess. target discovery is often difficult and may take years to complete . such as a virus or bacterium. and what options do they have for intervention? Since the human body is an extremely complex system. cancer. Why? Cells and cell-to-cell interactions are very complex . Different targets respond to different therapeutic approaches . chronic lower respiratory disease and diabetes .Initiating Drug Discovery Research An early step in the drug discovery process is to identify an unmet medical need . A target is a molecule that plays a critical role in a disease . The goal is to develop a drug that affects a target in a way that interferes with the disease process . disease processes take place at the molecular level . technology and financial resources to solve the problem? Potential competitors and barriers. BIoFaCT According to the National Center for Health Statistics. more complex chemicals or chemically related molecules from common chemical structures. often with improved product results or lessened product side effects. more-complex assays are used at subsequent levels of screening. protein electrophoresis.

cost and technology . the next step is to validate them .Cell Receptors and Ion Channels The most common drug targets are cell receptors—proteins on or inside a cell to which a specific signaling molecule can attach. What happens to those cells and tissues in the presence of a drug candidate? Does a drug candidate adversely affect other cells or tissues? Does it raise an immune response. or otherwise present any concerns about toxicity? BIoFaCT Recently. a next step is to use an animal model . The common thread among these targets is that they are often involved in signal transduction processes in and among cells . These signaling molecules can be hormones. pharmaceutical drugs. and only recently have mouse models been developed to mimic the disease . proteins that form pores in the membranes that surround cells. Sometimes the drug candidate is so specific to humans. toxins or even infectious agents. Other common drug targets are ion channels. Sometimes people who are born without certain functional molecules express a specific disease type . a physical change occurs that initiates a specific cellular response. Examples of target molecules include receptors. and enzymes— proteins that increase the rate of specific chemical reactions. Scientists also look at what other effects the drug candidate may have within preclinical (both cell culture and animal) models . There are a number of ways to validate a target. the concept of target validation is to use the target to create the disease in a sample of healthy tissues and then block the target to restore the healthy condition . The complexity of the body’s response also means scientists could see a difference in the expression of hundreds of genes without being able to determine which ones were critical to the disease . The trick is to select a model that is representative and will work . Initial studies are often done in cell culture . Sometimes the target doesn’t exist in an animal model or may not mimic the human disease state . ion channels. scientists have begun using computer simulation to model drug-target interactions to guide drug discovery . At the simplest level. If cell culture studies are positive. protein synthesis and programmed cell death (apoptosis). Target validation has two components . Signal transduction pathways control cellular processes such as division. differentiation. growth factors. For example. neurotransmitters. Alzheimer’s disease occurs only in humans. Preclinical work helps support later human trials that may occur if the drug candidate continues to show promise . When signaling molecules attach to the receptor. it won’t recognize the animal model’s target or the animal will mount an immune response that blocks any therapeutic effect . and the process must take into consideration time. The first is to show that the target molecule actually plays a role in the disease . stimulate other. Target Validation Once scientists identify potential targets. Sometimes a suitable animal model has to be created to validate a target . This is done in cell culture or animal models . Even if the drug gets marketing approval after successfully completing the necessary phases of human 20 Drug Discovery . Studying biological samples taken from such human subjects provides another means of validating a target . cytokines and DNA binding proteins . The second is to confirm the target is a candidate for therapeutic intervention: Can a safe and effective drug be made against the target? Scientists complete this second component of target validation before the drug enters human testing . enzymes. Sometimes the target is expressed on other cells or tissues besides those directly involved in the disease . similar targets.

partly because they are modeled on real biological molecules. Large batches of potential drugs are tested for binding activity or biological activity against target molecules . This means patients don’t have to take as many doses of a biologic. a company’s research lab develops a testing method (assay) to determine or measure the pharmacological activity of hundreds to hundreds of thousands of molecules . Each has its particular advantages and disadvantages. meaning. What’s being measured could be as simple as the ability of the drug candidate to kill cancer cells in culture or as complex as measuring its ability to inhibit an enzyme involved in a disease . antibodies or genes that modulate a particular biomolecular pathway . allowing them to be used for targets inside cells. Biologics often have much longer half-lives. Biologics usually need to be injected. the more complex the assay. Screening High-throughput screening is a process that combines robotics and data processing to rapidly identify the compounds. restricting their use to targets on the surface of. which may result in better patient adherence to therapy. Of the molecules that score a hit—that is. When designing a drug candidate. If the target is on the exterior surface of the cell membrane or is secreted. 3-D structural information about a target enhances drug design strategies . a positive result that appears to have a therapeutic potential—some are identified as lead molecules due to their more druglike properties (solubility. but recombinant antibodies generally have extremely high specificity. but biologics usually cannot. scientists may attempt to optimize its ability to fight disease by changing its molecular structure through combinatorial chemistry for small molecules or protein engineering for large molecules . Small molecules can usually cross cell membranes and enter cells. Considerations in designing a therapeutic agent depend on both the nature of the target and the capabilities of the company . Once a candidate disease is identified. cells. permeability. stability. scientists seek to develop a drug that is highly specific to a particular target in a disease in hopes of achieving a better therapeutic outcome with potentially fewer side effects . etc . safety surveillance will continue once the drug has reached the larger patient population . fewer adverse reactions for the patient. scientists must keep in mind the intended method of drug delivery and determine whether the drug will be a pill swallowed. Choosing the Right Tool for the Target Designing a targeting strategy usually comes down to a choice between a small-molecule drug and a biologic (most often a recombinant protein or antibody). whereas small molecules can be taken orally. Once a drug candidate is identified. Generally. Scientists will continue to answer safety questions throughout the life of a drug . which to date has limited their usefulness for treating diseases of the brain such as psychiatric disorders and neurodegenerative diseases. protein therapeutics such as monoclonal antibodies or peptides can be used . a liquid injected.) . Scientists can learn more about the structure of the target by using imaging technology such as X-ray crystallography . The assay measures the estimated potential of a molecule to block or stimulate a target . Drug Design The design approach to drug discovery starts with scientists understanding the genetic and molecular base of a disease and using that information to select a specific therapeutic target . Small molecules can often cross the blood-brain barrier. only drugs that can cross the cell membrane. Drugs are then designed to interact with the target . such as small molecules. Biologics usually cannot cross cell membranes. a spray inhaled or something else . Through rational drug design. Small molecules have variable half-lives. If the target is on the interior of the cell. or outside.trials. can be used . which is a measurement of how long a drug stays active in the bloodstream or in its target tissues. Drug Discovery Drug Discovery 21 . Small molecules have good specificity for their targets. the more relevant the information— but the higher the cost of the assay and the longer it usually takes to get data .

Drug development includes the safety. testing begins in humans in a series of studies called clinical trials. safety testing begins with a series of experiments called preclinical studies. Typically. the process of drug development is still far from complete. formulation and manufacture of the drug. If these studies predict the drug candidate to be safe.Chapter Six: Drug Development After the lengthy process of drug discovery (identifying a target and validating a drug candidate). 22 Drug Development . efficacy.

Knock-out mice are genetically altered to remove mouse versions of human disease genes . enabling researchers to check for adverse side effects before giving the candidate drug to humans . In target validation. visit www . research involving animals and that receive federal funding must have an Institutional Animal Care and Use Committee (IACUC) .* *For more information on Amgen’s commitment to ethical use of animals in research. what the drug candidate does to the body (pharmacodynamics). Pharmacokinetic and pharmacodynamic studies are used together to reach the goal of preclinical studies. which then answer the question: Is the drug safe? In the United States. Cell lines are engineered to express genes that are often responsible for adverse reactions . Human disease genes can also be knocked in to create mouse models with human diseases like cancer. In the United States. many studies of drug safety and toxicity are done using cell lines. is used to answer the question: Is the drug harmful or toxic to cells or organ systems? Toxicology studies address the potential of the drug or its metabolites to kill or damage cells and organs. Drug candidates are tested on these mice. The creation of cell line models has decreased the number of animals needed for testing (reducing cost and time) and helps accelerate the drug development process . and how fast does this occur? How is the drug or its metabolites (breakdown products) eliminated from the body? Pharmacodynamic studies examine dose-response effects and often monitor biochemical and physiological changes (such as enzyme activities. The goal of preclinical studies is to predict what the body does to the drug candidate (pharmacokinetics). which shows what the body does in response to the drug. Information from these studies is vital .amgen . Alzheimer’s and Parkinson’s . and whether the drug candidate may pose potential health hazards or toxic side effects . institutes that conduct Pharmacokinetic testing provides data to answer questions such as: How is the drug absorbed and transported? Which cells and organs are affected? What enzymes in the body break down the drug.html . cause cancer or cause reproductive problems. Animal models greatly enhance scientists’ ability to test the effectiveness and safety of new drug candidates . Pharmacodynamic testing. Many other countries follow global harmonized regulatory guidelines as well . including birth defects or sterility . Although drug companies are required to submit animal model data to regulatory agencies as part of the drug approval process.com/science/ethical_research .Preclinical Studies Preclinical studies are tests that take place in a scientifically controlled setting using cell cultures and animals as models . This committee reviews research protocols and evaluates the care laboratory animals receive . researchers may use knock-out mice and/or knock-in mice to validate a target . blood pressure and body temperature) in the test subject . heart rate. Drug Development 23 . preclinical studies must be conducted under FDA guidelines known as current Good Laboratory Practice . Initially. companies are taking steps to reduce the number of animals used in testing because of ethical concerns and the cost associated with facilities . diabetes. It allows researchers to estimate a safe dosage level for humans in phase 1 clinical trials . The IACUC is responsible for making sure labs comply with the Animal Welfare Act .

SAD: Single-Ascending-Dose Studies A few volunteers are given a small dose of the investigational new drug and observed. tolerability and safe dosage range . If unsuccessful. companies can begin phased clinical trials in humans . Samples of body fluids are collected with each increase in dosage level to understand how the body processes the investigational new drug. which protects the rights and ensures the safety of human test subjects and follows the U .S . BIoFaCT A crucial component of initiating a clinical trial is recruiting study subjects who agree to participate and sign a document called informed consent . particularly when testing oncology therapeutics . 2 and 3—and test progressively larger numbers of humans in each phase . Code of Human Research Ethics . A patient volunteer is someone who has the disease the drug is intended to treat . These are usually healthy volunteers who do not have a disease . proper dosage. It takes several years to complete all three clinical trial phases . clinical trials are halted. After the FDA approves the IND. Participants can withdraw their informed consent at any time . Each phase has a different purpose and helps researchers answer different questions . Clinical trials are conducted in three successive phases—1. The goal is to evaluate the drug’s safety. However. another group is given a slightly higher dose. MAD: Multiple-Ascending-Dose Studies The same volunteers receive higher and higher doses of the investigational new drug until the dosage reaches a certain level. Phase 1 Phase 1 trials represent the first time an investigational new drug is tested on humans . The testing group is often small. Clinical Trials Clinical trials are tests designed to determine the safety. Clinical trials are often managed by a contract research organization (CRO).How Is Dosage Determined? There are two types of phase 1 dosage studies: SAD studies and MAD studies. or the drug’s potential side effects are too risky to involve healthy subjects (such as using some chemotherapeutic agents) . companies submit an Investigational New Drug (IND) application to the FDA . Usually these patients have been unsuccessful with available treatments or have few treatment options. Potential subjects must be informed about all aspects of the study before they decide to participate . the drug candidate proceeds to the next phase . Some companies divide phase 2 trials into phase 2A (to assess dosage) and phase 2B (to assess efficacy) . Clinical trials are conducted at different testing sites . which is an independent organization . Phase 2 The goal of phase 2 trials is to determine the efficacy and safety of the investigational new drug among a larger group of patient volunteers—usually 100 to 300 people . 24 Drug Development . sometimes patient volunteers will be accepted into a phase 1 clinical trial. If a phase is successful. The CRO is responsible for all the data management and communication between the sponsor company and physicians overseeing the clinical trials . Most investigational new drugs fail during this stage because of efficacy and/or safety issues . Phase 1 clinical trials are usually conducted in an inpatient clinic where full-time staff can observe the study subjects. the investigational new drug is said to have reached maximum tolerated dose (MTD). efficacy. such as the FDA’s current Good Clinical Practice (cGCP). At these dosage levels. the drug is suspended and the sponsor company returns to the discovery phase to look for another drug candidate . If there are no adverse reactions. Clinical trials taking place in humans are conducted under global harmonized guidelines. This is repeated as many times as needed until volunteers start to exhibit intolerable side effects. If preclinical trials provide sufficient evidence that a drug candidate is safe. adverse reactions and long-term-use effects of a new drug in human subjects . The CRO also ensures that the study volunteers understand and accept the risks involved in the clinical trials and that cGCP guidelines are followed . ranging from 20 to 50 volunteers .

the sponsor company is permitted to market and sell the product in the country or countries regulated by that authority . If an adverse reaction is discovered. such as the FDA’s current Good Manufacturing Practice (cGMP). A goal is to monitor the drug’s safety and efficacy when utilized in a normal medical setting in a population of patients Phase 3 The goal of phase 3 trials is to confirm the effectiveness of the investigational new drug and compare it with placebos or therapies already available on the market . In this type of study.must take place in a facility that meets the country’s strict guidelines. The stages in product development. Most investigational drugs do not make it . After further testing. hundreds or thousands of patient volunteers are tested . Either the sponsor company can voluntarily withdraw the drug or a regulatory body can pull the drug from the marketplace . Sometimes adverse reactions. known as the EMEA. which were not seen in a comparatively small cohort of patients (3. take. Out of every 1. To do this. Study Designs Late-stage trials often include a double-blind randomized controlled test. neither the patient volunteer nor the researcher knows which volunteer belongs to the control group or the experimental group. to ensure safety and purity of the product . In the United States. lasting for a couple of years or longer to establish long-term safety . A third party keeps this documentation and releases it only after the study is over.000 potential new drugs in discovery. the drug may be withdrawn from the market . the company would file a New Drug Application (NDA) for a small-molecule drug or a Biologic License Application (BLA) for a large-molecule drug with the FDA . the drug may or may not be reinstated . in Europe) approves the drug.000 patient volunteers as compared to millions). Drug Development 25 . the sponsor company files a new drug or biologics application with the country’s regulatory agency . Once phase 3 is successfully complete. BIoFaCT One of the largest challenges associated with clinical trials is the shortage of study subjects . only one will make it to approval . on average. If the governing regulatory authority (the FDA in the United States or the European Medicines Agency. The final manufacturing of the drug—or large-scale production— that could number in the millions . Phase 3 trials are the most expensive and time-consuming. Each patient volunteer is randomly placed into one of the groups. Phase 4 Phase 4 trials occur after an approved drug is on the market . are discovered in larger and more diverse populations . or product pipeline. 10 to 15 years to complete .

and preparing the biologic for patients. 26 Scale-Up and Manufacturing .Chapter Seven: Scale-Up and Manufacturing The manufacturing of biologics is complex. The whole process from creating the master cell bank to preparing the biologic for patients can take years and cost hundreds of millions of dollars. The manufacturing of biologics has become a science that can be summarized in four key steps: producing the master cell line. since most are proteins—large molecules often variable in structure and sensitive to environmental conditions. isolating and purifying protein from cells. growing cells and producing protein.

nonsecreting (NS0) cells (pronounced “NS zero”) and E. meaning. Scale-Up and Manufacturing 27 . companies store their cell banks in two or more locations within their facilities and in one location off-site.000 liters or more There are a number of reasons to use these cells . Scale-Up Process The scale-up of a cell culture process can be very difficult and time-consuming. The media provides the nutrients and the optimum environment for cells to survive . The entire process of producing a biotech product from start to finish is often called a campaign and is usually divided into two main parts: upstream and downstream.Using R&D Specifications During the R&D phase. for clinical trials—for a biotechnology medicine. scientists remove and thaw a vial of cells from the cell bank and initiate a cell culture in a flask containing a small volume of growth media . To begin a campaign.atcc . insect or mammalian) growing in culture . the properties of the cell and the regulatory requirements . Common Cell Lines Many biotechnology products are proteins that must be produced by cells grown in culture . purification. Both CHO and NS0 cells synthesize proteins much like human cells do . Growth vessels vary in size: • Flasks hold 5 mL • pinner flasks or roller bottles S hold 50 to 200 mL • ench top bioreactors hold B 5 to 20 liters • ilot scale bioreactors hold P 50 to 200 liters • roduction vessels hold P 20. Using all of the R&D data from these production steps. they should be able to grow and produce product forever . or physical form. The initial volume of media can be as little as 5 mL . researchers develop the initial production methods on a small scale . Scientists use one vial of cells from the MCB to create the WCB. They also determine the drug’s final formulation. Both cell lines have generally regarded as safe (GRAS) status for therapeutic protein production . Researchers are well versed in their optimal culture conditions . nonprofit resource dedicated to the collection. The MCB is a reserve of cells that scientists use only when absolutely necessary. Scale-up is done by gradually transferring the growing cells into successively larger growth vessels containing larger media volumes . Downstream processes include the recovery. The selection of the cell line depends on the expertise of the company. BIoFaCT The American Type Culture Collection (ATCC) is a private.org) Other cell lines can be used and may be more suitable . Chinese hamster ovary (CHO) cells. To protect the integrity of the cell lines. Both are immortal cell lines. Upstream Phase Upstream processing begins with the cells that scientists create or engineer to make the protein product . Upstream processes involve production of the protein product. especially monoclonal antibodies . but they do not make or secrete any of their own antibody protein . taking as long as several months before researchers can obtain a product . usually an injection or infusion . coli are cell lines used for production of biotherapeutics. Working from the same stock of cell line reduces the chance of mutations associated with serial cultures. (www . the WCB is used to produce batches of product in the scale-up process. The scale-up and manufacturing process must adhere to cGMP guidelines to ensure product safety and purity . The cells are constantly dividing as long as the growth environment remains favorable . NS0 cells have the additional advantage of being programmed to produce antibodies. it is cryopreserved: scientists freeze a large number of vials of the cells to create a cell bank. companies devise large-scale production methods to produce enough of the product for the intended market . Once the desired cell line is made. preservation and distribution of authentic cell lines and other biologic materials . Once established. Therefore. more and more cells are present Cell Banks Cell banks involve a two-tiered frozen cell banking system: a master cell bank (MCB) and a working cell bank (WCB). most often by using cells (microbial. formulation and packaging of the protein product .

The greater the number of cells. assures product quality and testing during the product development stages well before the product is at the stage of marketing. After harvesting the protein product. Any contamination of a culture ruins the entire batch of product and costs a company money and time . scientists measure cell viability and concentration. which then has to be purified away from the other components that were inside the cell . This is where scientists separate the protein from cellular debris . Researchers verify the isolation and purification of the protein product through confirmed testing protocols . Proteins found outside of the cell (extracellular proteins) can be easier to isolate . They do this manually in the initial scale-up steps to optimize growth parameters such as temperature. yeast or other microorganisms . The protein product is then formulated according to the R&D specifications and packaged for use by physicians and patients . Using the same assays or testing methods used in the initial R&D stages. Purification of protein mixtures by column chromatography separates proteins based on physical and chemical properties such as size. shape or charge (+ or –) . product concentration and product activity at each incremental scale-up stage for monitoring purposes . the protein product is isolated from the cells that produced it . Technicians follow very strict protocols for maintaining aseptic conditions at all times during the scale-up and manufacturing stages . Proteins found inside the cell (intracellular proteins) require special protocols to extract them for purification . Then they apply the protein solution to a series of chromatography columns to obtain a pure protein product . Lab technicians monitor and control the physical environment in which cell cultures grow . Scale-Up Monitoring The goal of the scale-up process is to grow cells as quickly as possible and to produce as much protein product as possible . ensuring that the scale-up and manufacturing processes meet certain standards . It is crucial during the scale-up. the The monitoring process is automated once the cell culture is large enough to be grown in bioreactors .with each step . the more protein product is generated . Downstream Phase In the downstream phase of manufacturing. Usually this involves bursting the cells open to release the protein product. The QC department next step is clarification. fermentation and manufacturing stages that technicians monitor and test the cultures for contamination by bacteria. 28 Scale-Up and Manufacturing . The QA department is usually responsible for meeting and reporting quality objectives . nutrient concentration and oxygen level . pH. Additional purification steps remove any residual DNA and deactivate any viral particles that may be present . Quality Control and Quality Assurance Quality control (QC) and quality assurance (QA) departments are responsible for all of the monitoring that is crucial to the success of the scale-up and manufacturing stages of product development .

Chapter Eight: Biotechnology Medicines The biotechnology industry uses advanced technologies to apply cellular and molecular biology to create new. Medical biotechnology products are used to treat or prevent diseases. vaccines. blood components and tissues and cells for transplantation. These products include therapeutic proteins. Biotechnology Medicines 29 . monoclonal antibodies. beneficial products. allergy immunotherapy products.

In fact. and white blood cells will attack them . it took 20 years before the technology showed its true potential. attacked by a pathogenic virus . immunology.Monoclonal Antibodies Though monoclonal antibody technology was invented in the mid-1970s. and bind to human targets . Scientists use genetic engineering to create recombinant vaccines by inserting genes for desired antigens into a vector . do not cause disease but do have the antigen. thus tricking the body into thinking it is being Peptibodies are engineered therapeutic fusion proteins with attributes of both peptides and antibodies but are distinct from each. Doctors have long used therapeutic proteins to replace or supplement patients’ natural body proteins—especially when natural protein levels are decreased or lost due to disease . including tests for hepatitis and AIDS . Vaccines are also created with recombinant proteins . The first experimental monoclonal antibodies developed in mouse models were ineffective because the human immune system rejected mouse antibodies as foreign. Typically. Humanized antibodies are engineered to be mostly human to avoid problems with rejection . Antibodies can attach to antigens found on a pathogen and flag the pathogen for destruction by the immune system . scientists commonly use recombinant protein antigens as diagnostic reagents in enzyme-linked immunosorbent assays (ELISAs) for the detection of infectious agents such as Severe Acute Respiratory Syndrome (SARS) . antibodies has enabled the successful use of this breakthrough technology in fighting cancer and other serious illnesses. The first vaccines were made with inactivated (killed) or weakened virus unable to reproduce in the body but sufficient to provide immunity upon future exposure to the live virus . the body recognizes antigens as foreign. and other versions are not exact versions but produce similar effects in the body . Some biologics have been in use for more than 20 years and are considered standard therapy . then fully human. Fully human antibodies are derived from human cells or human antibody genes . Peptibodies Some recombinant proteins are versions of natural body proteins. scientists use recombinant DNA technology to produce a number of diagnostic tests for diseases. endocrinology and virology . Antibodies A major area of biologics is the production of humanized or fully human antibodies . Biologics are used in such fields as oncology. Recombinant vaccines. however. is a weakened virus or bacterium into which harmless genetic material from another disease-causing organism can be inserted . Approximately 50 recombinant therapeutic proteins are approved for clinical use and are currently marketed. or carrier. Recombinant vaccines are safe and easily grown and stored . The subsequent development of first humanized. 30 Biotechnology Medicines . often referred to as biologics . A vaccine vector. Diagnostics In addition to recombinant proteins being used as biologic drugs. Therapeutic Proteins Scientists use recombinant DNA technology to make therapeutic proteins. rheumatology. Vaccines Vaccines stimulate the immune system and provide protection against particular diseases . and hundreds more are undergoing clinical trials . Antibodies also can attach to proteins on immune cells that are involved in autoimmune responses in diseases like rheumatoid arthritis and multiple sclerosis .

innovative diagnostics and therapies are changing how some human diseases are prevented and others are treated. Future of Biotechnology in Healthcare 31 .Chapter Nine: Future of Biotechnology in Healthcare Biotechnology can offer patients more and better healthcare choices. New. with novel medicines. diagnostics and technologies in development that hold great potential to improve patients’ lives. This monumental healthcare shift is in its early stages.

The discovery of singlenucleotide polymorphisms (SNPs)—singlenucleotide changes in the DNA sequence— was one of the major breakthroughs in genetic testing . variations of the genome among individuals and variations of the encoded proteins produced enables researchers to develop medicines that address the individual needs of each patient . Pharmacogenomics and personalized medicine promise to improve clinical trials for new drugs. Utilizing technology to detect SNPs allows for more-accurate diagnosis of genetic diseases and therefore facilitates treatment decisions . determine its course. When a SNP occurs in a gene sequence that encodes for a specific protein. weight. Pharmacogenomics A major movement in healthcare is pharmacogenomics. Genetic testing provides patients with both an understanding of possible risks for certain diseases and possible opportunities for prevention . Pharmacogenomics takes advantage of the fact that individuals have unique genomes representing their genetic makeup . By understanding a patient’s genetic makeup. including age. SNPs (pronounced “snips”) represent one of the most common forms of genetic variation among individuals .Personalized Medicine Personalized medicine is the concept that patients should be treated with therapies and medicines based specifically on each patient’s unique genetic makeup. Each genome is likely to react differently to a particular drug and dose amount . it may change that protein and cause a disease or increase a patient’s susceptibility to a disease . These advances allow for testing and identifying an individual’s unique genetic makeup and then comparing those differences with the population at large . for optimal results . advance screening technology for diseases and result in moreeffective individualized healthcare and advances in preventive medicine . genetics and environment . diet. The challenge is to identify which drug and which dose will work most optimally for each person or for groups of individuals who share similar genetics . a physician can better prescribe a drug and dose level that will optimally work to combat a particular disease . 32 Future of Biotechnology in Healthcare . the practice of medicine is based on standards of care that are determined by averaging responses across large groups of people . gender. Knowledge of the human genome. RNA or protein molecules to identify a disease. BIoFaCT Approximately 10 million SNPs have been identified in the human genome . Advances in DNA technology are the keys to both pharmacogenomics and personalized medicine . Personalized medicine is a new paradigm that proposes to manage a patient’s disease based on the individual patient’s specific characteristics. Currently. Genetic testing is beginning to allow the development of genomic personalized medicine—medical care based on a patient’s genotype or gene expression profile . height. Genetic Testing The biotechnology industry has brought about vast improvements in testing and diagnosis for genetic diseases . evaluate responses to therapy or predict individual predisposition to a disease . BIoFaCT Molecular diagnostic tests analyze DNA.

The differentiated cells are then surgically implanted . because of their small size. could be a renewable source of replacement cells and tissues used to treat a wide range of diseases . Future of Biotechnology in Healthcare 33 . Nanotechnology deals with the manipulation of molecules and structures on a nanometer (one-billionth of a meter) or atomic scale . Nanoparticles called buckyballs—uniquely shaped and constructed carbon molecules— are also showing potential for drug delivery to target molecules or cells .Gene Therapy Gene therapy is an emerging area of applied genetics that utilizes recombinant DNA techniques . the recombinant DNA molecules themselves are used for therapy . disease-free cells—hence the term regenerative medicine for this approach . One example of nanomedicine is the experimental use of nanoshells to selectively target and destroy cancer cells at the cellular level . focuses its efforts on patients with severe and life-threatening diseases who usually have few treatment options or who have failed all available therapies . They may make it possible to deliver drugs that do not dissolve in water . Also. gene therapy research has expanded greatly. Nanoshells have the ability to capture infrared light shown through the skin of a cancer patient and convert it to heat. The field. Scientists are studying gene therapies for a number of inherited human diseases involving defective genes . Cell therapies also could be developed in which undifferentiated stem cells may be implanted along with growth factors to guide their differentiation in the patient’s body . Scientists are working on nanoparticles to unclog blocked arteries . they allow more of the drug to be delivered per volume . replace diseased cells and reverse the effects of the disease . Nanotechnology Since the first clinical trial was initiated in 1990. still in experimental stages. Applying nanotechnology for the improvement of human health is called nanomedicine. The broad potential of an undifferentiated stem cell to make a variety of other cells is the focus of stem cell research . into the cells and tissues of patients to treat their diseases . with an increasing number of human trials . The idea is to replace them with new. which kills only the targeted cancer cells . The theory is that stem cells may then integrate into the diseased tissue. In this case. Nanoshells are nanoscopic metallic lenses that are selectively delivered to specific organs or tumors through the bloodstream . The aim is to replace the damaged cells with healthy. Stem cells eventually differentiate to form all of the different types of cells that make up the body . created by recombinant DNA technology. Gene therapy involves inserting genes. functional genes . The hope is that stem cells. Biotechnology nanomedicine harnesses living organisms and/or their components on a very small scale . which is still in experimental stages. Stem Cells Stem cells are unspecialized cells that can renew themselves indefinitely to produce more stem cells . directed to differentiate into specific cell types. involves growing stem cells in the lab and guiding them toward a desired cell type by adding different growth factors . Stem cell therapy. They can mature and develop specialized functions or differentiate under specific growth conditions .

Current research is investigating the use of microspheres to deliver anticancer drugs to active tumors and for use with anesthetics in pain management . Microsphere therapies are currently available for lung cancer and respiratory illnesses . 34 Future of Biotechnology in Healthcare . They are made out of materials that resemble naturally occurring fats in cell membranes and are delivered as a mist sprayed into the nose or mouth .New Drug Delivery Systems Biomedical researchers are studying new ways of delivering drugs within the body that could improve effectiveness . One example is the development of microscopic particles called microspheres that have tiny holes just large enough to carry and deliver drugs to their targets .

and millions of patients around the globe continue to benefit from the treatments developed by companies that are discovering.Looking Ahead The practice of medicine has changed dramatically over the years through pioneering advances in biotechnology research and innovation. Future of Biotechnology in Healthcare 35 . future innovations in biotechnology research will bring exciting new advances to help millions more people worldwide. As companies continue to develop medicines that address significant unmet needs. developing and delivering innovative medicines to treat grievous illnesses.

” 36 Glossary . to make or modify products or processes for specific use . not normally present in the body that when introduced to the body stimulates a specific immune response and the production of antibodies . Angiogenesis is a key area of cancer research . Angiogenesis: The process by which the body forms and develops new blood vessels . computational especially when used in agriculture. it also allows malignant tumors to increase in size . prevention or treatment of disease . drug shelf life. Biologic: A product derived from a living organism (from animal products or other biological sources) that is used in the diagnosis. pathogenic processes or pharmacologic response to a specific therapy . It is a characteristic that is objectively measured and evaluated as an indicator of normal biologic processes. Biopharmaceutical: A synthetic drug produced utilizing certain biotechnology methods . Aseptic: Describes a product or method free a biologic state . Angiogenesis can be both beneficial and harmful.Glossary Amino Acids: The building blocks of proteins . The process of fermentation is performed in a bioreactor to grow large volumes of cells for producing specific proteins . manufacturing protocols. almost always a with the FDA seeking approval to market a novel biologic in the United States . The United Nations Convention on Biological Diversity defines biotechnology as “any technological application that uses biological systems. Apoptosis is a key area of cancer research . Examples of biologics include recombinant proteins. The application contains a description of the trials and results. living organisms. Biomarker identification and measurement are regarded as key developments for the future of disease treatment . Antigen: Any substance. Programmed cell death involves a series of biochemical events leading to characteristic cell changes and death . Bioinformatics entails the creation and advancement of databases. Antibody: A component of the body’s immune and statistical techniques. Assay: A test procedure whereby a property or concentration of a substance is measured . adenine (A) forms a base pair with thymine (T). Biotechnology: Technology based on biology. vaccines and hematopoietic growth factors . The unique sequence of amino acids in a chain defines the character of a protein molecule . Biologic License Application: An application filed response . food science and medicine . or derivatives thereof. It neutralizes the antigen by binding to it . In DNA. algorithms. and theory to solve formal and practical problems arising from the management and analysis of biological data . Bioinformatics: The application of information technology to the field of molecular biology . Biomarkers are also used in drug discovery to determine whether a drug is effective in animal models and at what doses effectiveness is reached . it is secreted in response to an antigenic stimulus . Bioreactor: A device or system for growing cells of microbiological organisms that would lead to contamination . as does guanine (G) with cytosine (C) . Base Pairs: Two nucleotides on opposite or tissues in the context of cell culture . Biosensor: A device that combines a biological component with a physicochemical detector component to detect a pathogenic agent . Apoptosis: The process of programmed cell death that may occur in multicellular organisms . while it can be used to stimulate development in new blood vessels to fight clogged arteries. thymine is replaced by uracil (U) . etc . A Y-shaped protein. In RNA. dosage. complementary DNA or RNA strands that are connected via hydrogen bonds . formulation. allergy shots. Autoimmune Disorders: Diseases whereby an individual’s immune system mounts an attack on a portion of its own tissues . Tissues undergoing such an attack can be destroyed in the process . packaging information. Biomarker: A substance used as an indicator of protein.

Cell Culture Technology: The growing of cells mixtures of different molecules may be separated from each other . Column Chromatography: A type of chroma- tography that uses a column for containing and separating a mixture . enzyme. protein or antibody . Colony Hybridization: The screening of a library with a labeled probe (radioactive. it is sometimes possible to replace animal testing with cell testing when evaluating the safety and efficacy of medicines . Codon: A string of exactly three mRNA bases A collection of stored chemicals that may be used in high-throughput screening for drug development . Chromosomes are large enough to be seen under a microscope . Individual proteins are then separated using chromatography methods . With mammalian cell culture. etc .) to identify a specific sequence of DNA. processes used to create copies of DNA fragments (molecular cloning). Combinatorial Chemistry: A discipline in which a large number of new chemicals are created. WCBs house cells used in pharmaceutical production grown from those maintained in an MCB so that their stability and uniformity are well characterized . nutrition and gene expression. Clinical Trial: A type of research study that evaluates the safety and efficacy of new drugs. After the protein product is harvested. which may include removing intracellular proteins from cells. particularly expression of recombinant proteins . Glossary 37 . Cloning: The replication of a DNA sequence tion is living or dead . Chromatography: A process by which complex that code for a specific amino acid during translation of mRNA into DNA . Testing for cell viability usually involves looking at a sample cell population and staining the cells or applying chemicals . These tests are required by regulatory agencies as a precondition of regulatory clearance to market . Clarification: A step in the downstream phase outside of living organisms . Cell Bank: A facility where cell lines are kept frozen and stored for later use . though they remain alive . all cells other than germ cells usually contain 46 chromosomes: 22 pairs of autosomes and either a pair of X chromosomes (in females) or an X chromosome and a Y chromosome (in males) . After a certain number of population doublings. Cell banks include master cell banks (MCBs) and working cell banks (WCBs) . The larger the chemical library. bioluminescent. In humans. In each pair of chromosomes. cells usually stop dividing. MCBs house primary cell strains that are kept stored and not used for production purposes . the better the chance that high-throughput screening will find a hit (a potential drug candidate) . With the exception of some derived from tumors. first introduced in the 1960s .Blood-Brain Barrier: A physiological mechanism that alters the permeability of brain capillaries so that some substances. clarification steps separate the protein from cellular debris . RNA. This is accomplished by subjecting the mixture to many repeated partitionings between a flowing phase and a stationary phase . while other substances are allowed to enter freely . Chemical Library (or Compound Library): from one organism to create an exact genetic copy. An established or immortalized cell line has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification . are prevented from entering brain tissue. toxicity screening. compiled into a library and screened for potential therapeutic use . CHO cells are the most commonly used mammalian hosts for industrial production of protein therapeutics . one chromosome is inherited from the father and one from the mother . such as certain drugs or toxins. most primary cell cultures have limited life spans . medical devices and biologics in human subjects . CHO cells are used in studies of genetics. Cell Lines: Generations of cells grown from original primary cells . Primary cells are cultured directly from a living organism . It is a commonly used method of purifying proteins . Chromosome: A threadlike linear strand of DNA and proteins in a cell that houses genes . cells (cell cloning) or organisms . Cell Viability: Determining whether a cell popula- of manufacturing a biologic . Chinese Hamster Ovary Cells (CHO cells): A cell line often used in biological and medical research.

Genetic mutations that lead to EGFR overexpression or overactivity have been associated with a number of cancers . Enzymes are the mediators of cell metabolism . Hematopoietic Growth Factors: Protein hormones produced by the body to regulate blood development. DNA Ligase: The enzyme that creates a bond between the ends of single-stranded DNA segments . High-Throughput Screening: The process of screening a sample of compounds rapidly and in parallel. is effectively stopped . then analyzing the results and choosing further screening compounds based on this information . Molecular systems interpret the sequence of these nucleic acids to produce proteins . cells by using enzymes to effect chemical changes . then separate the target molecules . Enzyme-Linked Immunosorbent Assay (ELISA): A designation that a substance is considered safe by experts under the conditions of its intended use . a functional or structural RNA molecule . Examples are CHO and NS0 cell lines that have GRAS status for therapeutic protein production . DNA Polymerase: An enzyme that attaches separation of DNA. Downstream Phase: Involves manufacturing processes including the recovery. Extracellular Proteins: Proteins found outside of a cell . formulation and packaging of the protein .Cryopreservation: A process whereby cells or whole tissues are preserved by cooling to low subzero temperatures . DNA ligase is the glue . Translation of this fusion gene results in a single new protein with functional properties derived from each of the original proteins . DNA Fingerprinting: A technique used to distinguish between individuals of the same species using only samples of their DNA . DNase (Deoxyribonuclease): Any enzyme that catalyzes the breaking up of linkages in the DNA molecule backbone . charge and/or shape . or culturing. usually affecting growth or metabolism . affecting the production and maturation of blood-forming cells . for example. Enzymes: The many proteins produced by charide units are added to proteins . Hormones: Substances produced by one tissue and conveyed to another through the bloodstream. DNA (Deoxyribonucleic Acid): DNA is a nucleic Fusion Proteins (or Chimeric Proteins): Proteins created through the joining of two or more genes that were originally coded for separate proteins . Glycosylation: The process by which oligosac- A biochemical technique to detect the presence of an antibody or an antigen in a sample . purification. Gene: A length of DNA that codes for a particular protein or. Placing the molecules in wells in the gel and applying an electric current moves the molecules through the matrix at different rates based on their size. any biological activity. The gel is the medium used to contain. Where restriction enzymes are the scissors of recombinant DNA technology. Generally Regarded as Safe (GRAS): complementary nucleotides to a single stranded human DNA . to make them capable of making new substances or performing new functions . A cell-surface receptor that is activated when bound by epidermal growth factor . Gel Electrophoresis: A technique used for the acid that contains the genetic information used in the development and functioning of all organisms . 38 Glossary . At these low temperatures. Genetic Engineering: Alteration of the genetic material of cells or organisms in order. Fermentation: A process of growing. It is commonly used to detect infectious agents . in certain cases. Electrophoresis refers to the use of electricity to move the molecules through the gel matrix . including the biochemical reactions that would lead to cell death. Half-life: A measurement of the time it takes for organisms to act as biochemical catalysts . RNA or protein molecules by using an electric current applied to a gel matrix . Epidermal Growth Factor Receptor (EGFR): a drug to lose half of its pharmacologic activity or half of its administered amount in the bloodstream or in its target tissues .

in a controlled environment such as in a cell culture or in cells grown in a petri dish . Interferon: A naturally occurring cell-signaling by cells that are all derived from a single antibody-producing cell . Monoclonal Antibody: An antibody produced system to achieve a therapeutic goal . Monoclonal antibodies are a type of immunotherapy . Cell cultures are used to express this recombinant DNA and produce these partial-mouse and mostly human antibodies . Non Secreting (NS0) Cells: Mouse myeloma cells that are used frequently in the production of recombinant antibodies . one phosphate and one base . Nucleus: The organelle within a living cell that contains genetic material and controls life functions . they are called monoclonal and can be used to continuously produce identical antibody molecules with these same therapeutic characteristics . Since the cells are all identical and are produced by cloning one specific cell in great numbers.Humanized Antibodies: Monoclonal antibodies that have been synthesized by using recombinant DNA technology to avoid the clinical problem of an immune response to foreign substances . Microarray: A tool that enables analysis of the produce a desired antibody in large amounts . Pathogen: A disease-causing agent such as a bacterium or virus . Ion channels are involved in a wide variety of biological processes and are a favorite target in the search for new drugs . Mass Spectrometry (MS): An analytic technique of the DNA double helix and RNA . Nucleotide: The name given to an individual unit ment outside of a living organism. Pegylation: The process of adding polyethylene for determination of the elemental composition of a sample or molecule . Neurotransmitters: Chemicals that are used to relay. It is also used for de- glycol to a therapeutic protein. A nucleotide contains one sugar. Hybridoma: A cell that has been engineered to termining the chemical structures of molecules. Immunotherapy: Modulation of the immune levels of expression of genes in an organism or comparison of gene-expression levels . Media: Nutrient-rich substances in which cells are grown . Humanized antibodies are produced by merging the DNA that encodes the binding portion of a monoclonal mouse antibody with human antibody-producing DNA . Hybridization: The process of joining two complementary strands of DNA or one each of DNA and RNA to form a double-stranded molecule . Nanotechnology: The study and creation of protein produced by the immune system in response to infections such as viral infections or parasites . amplify and modulate signals between a neuron and another cell . Once a cell capable of generating an antibody with desired therapeutic characteristics is selected. then measuring their mass-to-charge ratios . Nanomedicine: The medical application of nanotechnology . Investigational New Drug: A drug that has been approved by the FDA for use in human clinical trials . Intracellular Proteins: Proteins found inside a cell . Immortal Cell Line: An established cell line that has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification . Ion Channels: Pore-forming proteins that help establish and control the small voltage gradient across the plasma membrane of all living cells . which enables Glossary 39 . such as peptides or proteins . Hybridomas are created by fusing immortal tumor cells with antibody-producing B-lymphocyte cells that continuously synthesize identical (or monoclonal) antibodies . In vitro: The technique of performing an experi- systems and devices at the level of molecules and atoms . Messenger RNA (mRNA): A polynucleotide copy of a DNA gene that communicates the code for building a protein to ribosomes so that new proteins can be built . laboratory processes are used to clone (make large numbers of) these cells . MS consists of ionizing chemical compounds to generate charged molecules or molecule fragments.

Where DNA ligase is the glue of recombinant DNA technology. the receptor goes into a conformational change. Preclinical Trials (or Studies): Tests that take place in a scientifically controlled setting using cell culture and/or animals as disease models . a hormone. a pharmaceutical drug or a toxin. are long chains of amino acids . A protein is a long chain of amino acids joined together with peptide bonds and therefore is sometimes referred to as a polypeptide . to cut DNA at a certain location . Restriction Enzymes: Enzymes having the ability for creating millions of copies of a particular segment of DNA . Regenerative Medicine: Research into treatments that restore damaged cells with healthy. Proteins: Compounds (chains of amino acids) proteins with attributes of both peptides and antibodies but that are distinct from each and that can bind to human drug targets . Recombinant DNA: A form of DNA that does not exist naturally and is created by combining DNA sequences that would not normally occur together . If a scientist needs to detect the presence of a very small amount of a particular DNA sequence. which usually initiates a cellular response .” Protein Engineering: A process for isolating and studying proteins and generating proteins with 40 Glossary . The enzyme then performs a complementary template of the cDNA strand such that a double-stranded DNA molecule is formed . Poly- constituting the ultimate expression product of a gene . Peptide Bond: A bond that links together two or more amino acids .the therapeutic protein to stay in the body longer . Peptides: Short chains of amino acids . Proteomics: The study of proteins . Scientists use these enzymes to isolate certain types of DNA and place them into new environments . PCR can be used to amplify the amount of that sequence until there are enough copies available to be detected . restriction enzymes are the scissors . Polymerase Chain Reaction (PCR): A method recombinant DNA technology . peptides. proteins are the workhorses of living systems. or multiple peptides linked together by peptide bonds. Proteomics has three major goals: to identify and quantify all the proteins expressed in an organism. Receptor (Cell Receptor): A protein molecule. Product Pipeline: In the biomedical industry. causing chemical processes and changing as their environment changes . Created through the synthesis performed by ribosomes. Personalized Medicine: Use of the information contained within a patient’s genome. Pharmacodynamics: Studies performed to determine what a drug does to the body . Recombinant Proteins: Proteins created by mine what the body does to a drug . and when such binding occurs. Reverse Transcriptase: An enzyme used by retroviruses to form a complementary DNA sequence (cDNA) from an RNA template—usually the genome of the retrovirus . Drugs that have entered into clinical trials are said to be “in the pipeline . and may be a peptide (such as a neurotransmitter). to which a mobile signaling (or signal) molecule may attach . Phosphorylation: The addition of a phosphate (PO4 ) group to a protein or other organic molecule . A molecule that binds to a receptor is called a ligand. Pharmacokinetics: Studies performed to deter- embedded in either the plasma membrane or the cytoplasm of a cell. to determine the structure and function of each protein and to study the protein-protein interactions that affects how one protein interacts with other proteins to control cellular processes . Pharmacogenomics: The science of under- standing the correlation between patients’ genetic makeup (genotype) and their responses to drug treatment . diseasefree cells . genotype or genomic signature to design and tailor the best treatment plan for that individual patient . Peptibodies: Engineered therapeutic fusion modified structures by altering the genes that direct their composition . Protein phosphorylation plays a significant role in a wide range of cellular processes . This double-stranded DNA molecule is then inserted into the chromosome the term pipeline refers to the number of unique products or processes reported or in development by a company .

Transgenic: A term describing an organism protein synthesis occurs . Upstream Phase: Involves the production of the protein product. Ribosome: The cell structures within which Transfer RNA: Molecules that carry amino acids during the process of protein synthesis during translation . From the angles and intensities of these scattered beams. The cycler then raises and lowers the temperature of the block in discrete. drugs. Scientists use this process to introduce recombinant DNA to bacteria. RNA is a nucleic acid transcribed from DNA. in which a beam of X-rays strikes a crystal and scatters in many different directions . ability both to multiply and to differentiate into specific cells . and the structure of a substance can be determined . preprogrammed steps . Vector: (1) An organism that serves to transfer a sequence variation that occurs when a single nucleotide—A. vaccines and other agents . human cells with the ing the arrangement of atoms within a crystal. mRNA is then translated into proteins . which has been infected by the retrovirus . Variations in the DNA sequences of humans can affect how humans develop diseases and respond to pathogens. Scientists are attempting to learn more about this process in cancer cells in order to fight the disease . Thermocycler: A laboratory apparatus. insect or mammalian) growing in culture . most often by using cells (microbial. which helps in the process of decoding the genetic information carried by DNA . Southern Blotting: Transfer by absorption of disease-causing organism (pathogen) from one organism to another . The device has a thermal block with holes where tubes holding the PCR mixtures can be inserted . T. This method has been referred to as posttranscriptional gene silencing and is an important tool for gene-expression research . used to amplify segments of DNA via the polymerase chain reaction (PCR) process . (2) A mechanism whereby foreign genes are moved into an organism and inserted into that organism’s genome . X-ray Crystallography: A method of determin- DNA fragments separated in electrophoretic gels to membrane filters for detection of specific base sequences by radiolabeled complementary probes . yeast and mammalian cell lines . Vaccine: An agent bearing antigens on its from the outside of a cell to the inside . G or C—in the genome differs between members of a species . Signal Transduction: The movement of signals sequence into a string of amino acids that form a protein . Translation: The process that converts an mRNA gene expression at the stage of translation (see translation) or by hindering the transcription (see transcription) of specific genes . Glossary 41 . a crystallographer can produce a three-dimensional picture of the density of electrons within the crystal. Ribonucleic Acid (RNA): A molecule similar to DNA. chemicals. Transformation: The process of transferring DNA from a donor to a recipient cell . Stem Cell: Undifferentiated. Translation follows transcription (see transcription).of the host cell. RNA Interference: A mechanism that inhibits containing genetic material from a source other than its parents . Transcription: The process by which enzymes use the genetic information on a strand of DNA to create a complementary strand of messenger RNA . Single-Nucleotide Polymorphism (SNP): A DNA surface that causes activation of the immune system without causing actual disease .

1964 .DNA ligase. Har Gobind Khorana synthesizes the first complete gene at the University of Wisconsin–Madison . . 1958 .The existence of reverse transcriptase is predicted . . 1953 . Joseph Murray performs the first kidney transplant between identical twins . Osamu Shimomura of Boston University discovers the green fluorescent protein Aequorea victoria in jellyfish . These enzymes cut DNA into pieces and are used for various studies and applications .Scientists understand genetic code for the first time . .The first automatic protein sequencer. the Moore-Stein amino acid analyzer.The first vaccine for rubella is developed . 1957 . containing HeLa cells. 1969 . Arthur Kornberg of Washington University in St . DNA polymerase. is isolated for the first time .Independent groups in the United States. 1970s 1970 . which links DNA fragments together. is used for the first time . Francis Crick reveal the 3-D structure of DNA .An enzyme. 1973 . 1972 .Dr . .Dr .Restriction enzymes are discovered . 1963 . The development marks the first use of mammalian cells (monkey kidney cells) and the first application of cell culture technology to generate a commercial product . 1967 . John F .Dr .Dr . is commonly used in medical research . Germany and China synthesize insulin.Dr . 1961 .The purified enzyme reverse transcriptase is first used to synthesize complementary DNA from purified messenger RNA in a test tube .Dr . Stanley Cohen and Dr . Jonas Salk develops the first polio vaccine . James Watson and Dr . is developed . which inserted 42 Timeline of Medical Biotechnology . He later develops . a pancreatic hormone . This cell line. Samuel Katz and Dr .Scientists prove that sickle-cell anemia occurs due to a change in a single amino acid .Timeline of Medical Biotechnology 1950s 1952 it into a tool for observing previously invisible cellular processes . who died of the cancer in 1951 . It is combined with the measles and mumps vaccines to form the measles/mumps/rubella vaccine in 1972 . Enders develop the first vaccine for measles . George Gey establishes a continuous cell line taken from a human cervical carcinoma isolated from Henrietta Lacks. involved in the synthesis of a nucleic acid. The restriction enzyme technique becomes a fundamental tool in modern genetic research and opens the way for gene cloning. . . 1960s 1960 .Dr .French scientists discover messenger RNA (mRNA) . Maurice Hilleman develops the first American vaccine for mumps .Dr . 1955 .Dr . Herbert Boyer use bacterial genes to perform the first successful recombinant DNA experiment.The DNA composition of humans is discovered to be 99 percent similar to that of chimpanzees and gorillas . Louis makes DNA in a test tube for the first time .Dr . . 1962 .

Dr . National Institutes of Health (NIH) forms a Recombinant DNA Advisory Committee to oversee recombinant genetic research .The U . San Francisco.The first vaccine for chicken pox is developed in Japan .Colony hybridization and Southern blotting are developed for detecting specific DNA sequences .S .Dr .a recombinant DNA molecule into a cell for replication .Protocols are developed to rapidly sequence long sections of DNA . 1982 . . marking the first time a synthetic recombinant gene is used to clone a protein .The first transgenic animals are produced by transferring genes from other animals into mice . .The first genetic markers for specific inherited diseases are found .Genetically engineered bacteria are used to synthesize the human growth protein somatostatin.Molecular hybridization is used for the prenatal diagnosis of alpha thalassemia .The FDA approves a monoclonal antibodybased diagnostic test to detect Chlamydia trachomatis. is developed in California .S . It becomes a seminal technology for studying the structure of DNA . Niels Jeme develop monoclonal antibody technology by fusing immortal tumor cells with antibody-producing B-lymphocyte cells to produce hybridomas that continuously synthesize identical (or monoclonal) antibodies . 1974 .Dr . .S . . 1980s 1980 .The NIH releases the first guidelines for recombinant DNA research . 1981 .The first monoclonal antibodies are produced . Luc Montagnier of the Pasteur Institute in Paris isolates the AIDS virus .The first vaccine for meningococcal meningitis is developed .Dr . 1976 . Supreme Court rules genetically altered life forms can be patented. opening up enormous possibilities for commercially exploiting genetic engineering . PCR is recognized as the most revolutionary molecular biology technique of the 1980s . 1975 . or genesynthesizing machine. Alaska . Edwin Southern develops a blotting technique for DNA called the Southern blot . .The first test-tube baby. . Dr . . patent for gene cloning . .The first artificial chromosome is synthesized .Scientists in Switzerland clone mice .The first automatic gene machine. 1983 . a technique for multiplying DNA sequences . César Milstein. . Food and Drug Administration (FDA) approves the first biologic—or recombinant protein: Humulin®. Robert Austrian of the University of Pennsylvania develops the first vaccine for pneumonia . . . which will grow to become the world’s largest biotechnology medicines company .The U .The U . Herbert Boyer of the University of California. 1978 .Dr . The first patent of this nature was awarded to the Exxon oil company to patent an oil-eating microorganism.Dr . Kary Banks Mullis invents the polymerase chain reaction (PCR). Baruch Blumberg and Dr .Dr . Herbert Boyer receive a U .Founding of Amgen. Georges Kohler and Dr . . which would later be used in the 1989 cleanup of the Exxon oil spill at Prince William Sound.S . Timeline of Medical Biotechnology 43 . constructs a synthetic version of the human insulin gene and inserts it into the bacterium E. Irving Millman develop the first vaccine for hepatitis B (not recombinant) four years after the virus is discovered . . 1977 .Yeast genes are expressed in E. Stanley Cohen and Dr . Louise Brown. . . . Genentech’s human insulin drug produced by genetically engineered bacteria for the treatment of diabetes . is born in the United Kingdom . . coli. . allowing the bacterium to produce human insulin .Dr . coli bacteria . Many consider this to be the advent of the Age of Biotechnology .

The FDA approves a genetically engineered tissue plasminogen activator to treat heart attacks . which are expression vectors for large proteins . .The first federally approved gene therapy treatment is performed successfully on a 4-year-old girl suffering from an immune disorder called adenosine deaminase deficiency . the use of a collection of distinct DNAs in arrays for expression profiling.The FDA approves a modified enzyme for enzyme replacement therapy to treat severe combined immunodeficiency disease . . In 1988.The FDA approves the first genetically engineered human vaccine to prevent hepatitis B . 1989 .The FDA approves first biotech-derived interferon drugs to treat cancer .The automated DNA sequencer is invented in California . . . 1985 chromosomes. Such variations in DNA are called restriction fragment length polymorphisms and are extremely useful in genetic studies . . .Genentech becomes the first biotechnology company to launch its own biopharmaceutical product . . the resulting sets of fragments sometimes differ markedly from one person to the next .The NIH approves guidelines for performing experiments in gene therapy on humans . It is the first successful application of enzyme replacement therapy for an inherited disease .DNA microarray technology. Maynard Olson and colleagues at Washington University invent yeast artificial .The FDA approves a bioengineered form of the protein interferon gamma to treat chronic granulomatous disease .The DNA fingerprinting technique is developed . This is a major step toward reducing premature birth complications .The FDA licenses the first hepatitis C antibody test. the international effort to map all of the genes in the human body. .Oil-eating bacteria are used to clean up the Exxon Valdez oil spill . .A gene responsible for cystic fibrosis is discovered . . chemist Dr . a massive effort to map and sequence the human genetic code as well as the genomes of other species . . Estimated cost: $13 billion .The FDA approves the first monoclonal antibody treatment to fight kidney transplant rejection . viruses and bacteria are field-tested for the first time . .The Human Genome Project. a complication of AIDS .Genetic fingerprinting enters the courtroom . which helps to ensure the purity of blood bank products . .The FDA approves Amgen’s first biologically derived human therapeutic .Dr .The first agreement between two companies with parallel patents for cross-licensing of biotech products occurs and becomes the prototype . When a restrictive enzyme is applied to DNA from different individuals. . . 1988 .1984 . .Genetically engineered plants resistant to insects.Reverse transcription and the polymerase chain reaction are combined to amplify messenger RNA sequences . 1986 . . The arrayed DNAs are used to identify genes whose expression is modulated by interferon .The FDA approves a diagnostic serum tumor marker test for ovarian cancer . .Cloning of the gene that encodes human lung surfactant protein is accomplished .Congress funds the Human Genome Project. . . Peter Schultz describes how to combine antibodies and enzymes (abzymes) to create therapeutics .The first genetically engineered vaccine is developed for hepatitis B . is first described . Berkeley. 1987 1990s 1990 . 44 Timeline of Medical Biotechnology .University of California. .The entire genome of the HIV virus is cloned and sequenced . is launched . . the drugs are used to treat Kaposi’s sarcoma.

. 1994 . Saccharomyces cerevisiae. coli.The U . are identified and their functions described .A new. . . produces a rough map of all 23 pairs of human chromosomes .The FDA approve the first genetically engineered blood-clotting factor—a recombinant protein used to treat hemophilia A .The FDA approves a recombinant protein to treat renal cell cancer . . inexpensive diagnostic biosensor test is developed to allow instantaneous detection of a toxic strain of E. hearing loss. cerulean cataracts. an international biotechnology advocacy group .The FDA approves the first bloodless HIV-antibody test that uses cells from patients’ gums .S .The FDA approves a recombinant protein to treat growth hormone deficiency . the bacteria responsible for many food-poisoning outbreaks . Daniel Cohen of the Center for the Study of Human Polymorphisms in Paris. .Scientists at the Institute for Genomic Research complete the first full gene sequence of a living organism (other than a virus) for the bacterium Haemophilus influenzae.Two smaller trade associations merge to form the Biotechnology Industry Organization. 1996 .The first human artificial chromosome is created .A group of scientists sequence the complete genome of a complex organism. A combination of natural and synthetic DNA is used to create a genetic cassette that can potentially be customized and used in gene therapy .The FDA approves a recombinant folliclestimulating hormone to treat infertility . BRCA1 . a gene associated with apoptosis B (programmed cell death) • edgehog genes (named because of their H shape) produce proteins that guide cell differentiation in advanced organisms • pr. Mary-Claire King at the University of California. thyroid cancer. a gene governing reproduction of the V HIV virus .Linkage studies identify genes for a variety of ailments. These include: • b. the U . human and otherwise. . . sudden infant death syndrome. .The FDA approves a recombinant protein to treat multiple sclerosis—marking the first new multiple sclerosis treatment in 20 years . which breaks down protein accumulation in the lungs of cystic fibrosis patients . . 1993 version of human DNase. Army and the Centers for Disease Control and Prevention are significantly involved in the development and clinical testing of the vaccine . . prostate cancer and dwarfism . led by Dr . a breast cancer susceptibility gene B • CL-2.The first baboon-to-human bone marrow transplant is performed on an AIDS patient . . . otherwise known as baker’s yeast . . including bipolar disorder. . . . a small glass or silica microchip that contains thousands of individual genes that can be analyzed simultaneously . 1995 . dyslexia.Dr . The NIH. discovers the first breast cancer gene.A multitude of genes. 1997 .A European research team identifies a genetic defect that appears to be the most common cause of deafness .The first vaccine for hepatitis A is developed . melanoma.Scientists at the Institute for Genomic Research sequence the complete genome of the Lyme Timeline of Medical Biotechnology 45 .American and British scientists unveil a technique for testing embryos in vitro for genetic abnormalities such as cystic fibrosis and hemophilia . It represents the first new therapeutic drug for managing cystic fibrosis in more than 30 years . The achievement marks the complete sequencing of the largest genome to date—more than 12 million base pairs of DNA . . a gene predisposing to obesity O • CR.The FDA approves a modified enzyme to treat Gaucher’s disease .S .An international research team. Berkeley.1992 . Army collects blood and tissue samples from all new recruits as part of a genetic dog tag program aimed at better identification of soldiers killed in combat .The Department of Biochemistry at Stanford University and Affymetrix develop the GeneChip. This marks a technological breakthrough in gene expression and DNA-sequencing technology .The FDA approves a genetically engineered .

2008 . 1999 . 2003 . dog and hundreds of other species .The FDA approves the first monoclonal antibody that is an antiangiogenic. . . showing the locations of more than 30. . . 2000s 2000 . This is an important step toward personalized medicine .Approval of the Her-2 inhibitor for the treatment of breast cancer patients who have tested positive for the Her-2 mutation brings personalized medicine to oncology . Parkinson’s and Alzheimer’s . .The FDA approves the first therapeutic antibody to treat cancer in the United States .Chemists in Japan create the first DNA molecule made almost entirely of artificial parts . inhibiting the growth of blood vessels—or angiogenesis— for cancer therapy .A rough draft of the human genome map is produced. Louis sequence the first complete animal genome for the Caenorhabditis elegans worm .Scientists at the University of Wisconsin– Madison sequence the E.Human skin is produced for the first time in the lab . . along with the genome for the organism linked to stomach ulcers. Embryonic stem cells are used to regenerate tissue and create disorders mimicking diseases . which causes genital warts and can cause cervical cancer .Two research teams culture embryonic stem cells . . making it possible for scientists all over the world to begin researching new treatments for diseases that have genetic origins. The discovery can be used in the fields of gene therapy .The FDA approves a novel monoclonal antibody to treat Crohn’s disease .A monoclonal antibody therapy used against breast cancer has favorable results.Scientists discover how to use human skin cells to create embryonic stem cells . Borrelia burgdorferi. 2006 . 2004 . It is used for patients with non-Hodgkin’s lymphoma . chimpanzee.000 genes .The FDA clears a DNA microarray test system. .disease pathogen. Craig Venter and his team replicate a bacterium’s genetic structure entirely from laboratory chemicals. Helicobacter pylori.An era of very rapid shotgun sequencing of major genomes is completed . 46 Timeline of Medical Biotechnology . .The complete genetic code of the human chromosome is deciphered . . 2007 . such as cancer. which aids in selecting medications for a variety of conditions . moving a step closer toward creating the world’s first living artificial organism .Celera and NIH complete sequencing of the human genome . 2001 .Scientists determine the 3-D structure of the human immunodeficiency virus.The FDA approves a recombinant vaccine against human papillomavirus.Science and Nature magazines publish the human genome sequence. Included are the mouse. heralding a new era of treatment based on molecular targeting of tumor cells . . coli genome . heart disease.The first vaccine for Lyme disease is developed . . .Scientists at Celera Genomics and the Human Genome Project complete a rough draft of the human genome . 1998 2002 .Dr . which causes AIDS .Scientists at the Sanger Institute of the Washington University School of Medicine in St .

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