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Volume 13, Issue No.

1

Planting
for the

Future
Plant-Made
Pharmaceuticals

................ Ed..... Muncy Junior/Senior High School Sharon Terry.. 12 Career Profile Anne-Marie Stomp ..... President Biotechnology Institute Contents Plants: Factories of the Future........ We focus on these basics: • What is involved in making plant-made pharmaceuticals (PMPs)? • How are PMPs different from nutraceuticals and edible vaccines? • Who can benefit from PMPs? • Are plant-made pharmaceuticals safe and effective? • How are policymakers dealing with PMPs? Biotechnology is shaping the lives of people of all ages............ .... Sc............. M... and Ortho Biotech.......................8 The Chosen Ones.... . Some back issues are available...............D...... Acknowledgments The Biotechnology Institute would like to thank the Pennsylvania Biotechnology Association...... North Montco Technical Career Center Anthony Guiseppi-Elie......... ...... ALL RIGHTS RESERVED...............to 12th-grade students.. Advisory Board Don DeRosa......org Phone: (703) 248-8681 Fax: (703) 248-8687 Biotechnology Institute The Biotechnology Institute is an independent..... founding editor....................D................. CityLab................15 Glossary and Resources ..... Baron Design Dodds Design Cover and Inside Illustration ©2003 John Michael Yanson ALL RIGHTS RESERVED. or library sets)... food.4 Worth the Effort ..... and educate the public. Published biannually......D............. Paul A............... Director of Education.............. teacher............. and Jeff Alan Davidson. Suite 202 Arlington........Main Points The Biotechnology Institute is pleased to present Your World’s fall 2003 issue exploring plant-made pharmaceuticals... about biotechnology and its immense potential for solving human health........... Genetic Alliance For more information Biotechnology Institute 1840 Wilson Boulevard... Your World is the premier biotechnology publication for 7th.. Hanle..................... Boston University Medical College Lori Dodson......... national........... the environment... ©2003 Biotechnology Institute. Our mission is to engage........... Ph...................... Issue No........ Mark Temons.... GeneData (USA).....A..... Interest in this field is blossoming as scientists recognize the tremendous potential of using plants to manufacture useful proteins........... Each issue provides an in-depth exploration of a particular biotechnology topic by looking at the science of biotechnology and its practical applications in health care............... nonprofit organization dedicated to education and research about the present and future impact of biotechnology............... Virginia Commonwealth University Lynn Jablonski...........................................14 Activity Microbial Bioassay..6 Down on the ‘Pharm’ . and environmental problems.. excite........D.... and industry... which originally developed Your World.....10 Potent Plants... It’s one of the many aspects of biotechnology—the use of living organisms to benefit humanity... agriculture..................................................................... and we hope this magazine cultivates an interest in a career that is part of this exciting technology... Inc... Troy Mashburn Photo ©2003 Keith Barraclough ALL RIGHTS RESERVED. Ph.......... 2 Planting for the Future: Plant-Made Pharmaceuticals ....2 So You Want to Make a PMP ........................ President............ particularly young people..... VA 22201 info@biotechinstitute........ The Biotechnology Institute acknowledges with deep gratitude the financial support of Centocor... Inc................................... Please contact the Biotechnology Institute for information on subscriptions (individual.........16 Volume 13..... 1 Fall 2003 Publisher The Biotechnology Institute Editor Kathy Frame Managing Editor Lois M..

the plant is merely used as a factory for protein production. exposure of farm workers to potentially harmful genetic material.m. There are legitimate concerns about using plants to produce therapeutic proteins. Plants are naturally good protein makers. as in “I’ve got to clock in at the plant at 5 a. The following articles give you details about this up-and-coming field. sunlight. Scientists are working hard to address these issues. coli/travelers’ disease and cystic fibrosis. Department of Agriculture (USDA) and the FDA oversee every stage of PMP production. air. Plant-made pharmaceuticals do not involve eating the plant. minerals. approved by the Food and Drug Administration (FDA) in the United States. The proteins for PMPs are always extracted from a plant. Some PMPs are already being tested in humans. These diseases could go from arthritis to asthma to cancer and beyond. protein production in bacterial systems and mammalian cell cultures is very costly and limited by the capacity of buildings and equipment. Clinical trials using proteins encoded in corn to treat E. At the very least. roots. and the right set of genes. and government agencies have set up strict rules and routines for confinement and cultivation methods that minimize risks. that is. and a tobacco-based PMP to treat non-Hodgkin’s lymphoma. Researchers also understand quite a bit about plant genetics. we may see plants—the kind with seeds. growth. PMPs are true drugs. vaccines administered by eating a specific amount of a food. They should not be confused with nutraceuticals. Your World 3 . which might provide more people with access to these drugs and vaccines. Using biotechnology. scientists place into a plant a gene that expresses a medically useful protein. the science promises a new ability to expand production. which are compounds used as part of a diet to improve health. These protein-based drugs and vaccines (“biologics”) are called plant-made pharmaceuticals (PMPs). Changing the balance of production cost and profit also may make it possible to offer drugs that are currently too expensive to produce in mass quantities. Scientists hope that several PMPs will be commonly available in three to five years. and development.” In the not-too-distant future. The U. Plants can be turned into economical and renewable protein-manufacturing systems with a few simple resources—water. and scientists know a lot about how plants go about their work. workers have been reporting to manufacturing facilities that they casually call plants. and the possibility that wildlife and insects will feed on the altered crops. are under way. In contrast. stems. which makes cultivating genetically modified plants relatively simple. This technology holds tremendous potential to expand the range and availability of pharmaceuticals to treat a wide array of diseases.S. and leaves—serving as factories for plant-made pharmaceuticals. including a low risk of food and feed contamination.Plants: Factories of the Future A cross the country. Plant-made pharmaceuticals could dramatically benefit anyone who uses drugs or hopes to find a drug to treat his or her illness. then the plant makes that protein using its own biological machinery. And they are not edible vaccines.

actually works pretty well! Unfortunately the results are not consistent (imagine the random results from shotgun pellets). •Grow the plant cells up into whole plants. for instance. it tells the RNA to stop reading.How are plant-made pharmaceuticals produced? So You Want to Make a S o how do you turn a plant into a pharmaceutical factory? In other words. so biolistics is not widely used. When the bacterium invades the plant. •Transfer the gene into plant cells grown in a dish. how can you separate that protein from all the other proteins the plant makes? Surprisingly. But a gene needs another DNA sequence near its beginning to tell where and when it should be “read” (or in scientific terms. fully developed plants. sending our protein to its destination within the cell. like peanut oil or olive oil. it is the site on the 4 Planting for the Future: Plant-Made Pharmaceuticals PMP. DNA where RNA polymerase will bind and begin transcription.. Such a sequence is called a promoter. and use a gun to shoot them into plant cells.) We should also add another bit of DNA called a signal sequence. More than two decades of research in genetic engineering— including the knowledge of how to move a gene from one living organism to another—has advanced these techniques from obscure art to routine science. Adding an oil body signal sequence to our gene will direct our protein here after it is produced. the plasmid is transferred into the plant’s own cells. this process. •Harvest the plants. 10. carrying the gene along with it. Believe it or not. A second method is to place a gene in a small circle of DNA (plasmid) that is located in the common soil bacterium. since the oil body serves as a foundation that protects the proteins. Where shall we send it? One option is the plant’s oil body. the essential tools are well known and widely available. Transferring a Gene into the Plant Two major methods are open to us.. Controlling a Gene A gene is the segment of DNA involved in making a protein. and isolate the protein. and let the bacterium infect a plant. And when it comes time to purify the protein from the plant. The first step in making any PMP protein is to isolate the gene for the protein we want. Let’s look at each step in more detail. We also add a termination sequence. the rest of the process looks like this: •Add other bits of DNA to help control the gene (to express the right protein) once it’s inside the plant. (See also “The Chosen Ones. Once we have the gene. where and when it should be transcribed). called biolistics. and it makes an ideal spot for proteins too. we can add the same promoter that plants use to turn on genes in their own seeds. If we want an antibody gene to be expressed only in seeds. The signal sequence DNA codes for a little tail of amino acids on our protein that acts like an address label.” p. Agrobacterium tumefaciens. how do you make a plant start manufacturing a protein it has never made before? And when it’s done. this isn’t quite as hard as it sounds. which we’ll tag onto the end of the gene. a membrane network within the cell. This compartment stores oil. While a great deal of skill (and patience!) is still required. the tools are even more well known—people have been extracting useful substances from plants for centuries. We can stick the gene we’re making onto microscopic particles of gold. it is much easier to infect . which is like a period at the end of a sentence. Other PMPs may be easier to isolate if they are stored in the endoplasmic reticulum (ER). Although it’s possible to infect mature.

how do we get the protein out? First we harvest the plant part— seeds. —Richard Robinson Your World 5 . How does a company keep someone else from planting and profiting from their invention? One strategy is to insert a "signature sequence" of DNA just after the signal sequence. discouraging theft. This puts the company’s unique "fingerprint" on the gene. Once the plants have taken up the gene. 10). A signature sequence also makes it traceable in cases of contamination. If we’ve routed the protein to the oil body in the seed. provide a highly controlled environment for manipulating plant cells. as they are called. they are ready to be transplanted into soil and grown in a greenhouse or a field. Before they begin a project. Fuel Protein separates the cell into different parts.Green Machines Chloroplasts are the green machines inside plant cells that convert sunlight. Affinity chromatography: The protein mix passes through a matrix—that is. Therapeutic Proteins PMP METHOD Gene Programming Disposal of Biomass Integration of Gene Into Plant Production of Plant Material (Biomass) Recovery and Quality Control Gene of Interest Energy TRADITIONAL MANUFACTURING METHOD Cell Culture thousands of small pieces of plant tissue growing in a lab dish. a column filled with an insoluble substance such as beads. we will probably grind the leaf up to release its contents. Such tissue cultures. such as endoplasmic reticulum or chloroplasts. from gene construction through final purification. we alter the mix of hormones in the growth medium to encourage development. Within several months. to get the most for the least cost. Here are some possibilities: Filtration: Removes plant fibers. and water into sugars. or roots—containing this protein.” p. The final wash releases the desired protein all nice and clean. but it does indicate the origin of the transgenic plant. air. This unique bit of DNA doesn’t make any protein. extraction will depend on the particular chemical properties of the protein we’re interested in and the tissue it’s in (described in "The Chosen Ones. Ultracentrifugation: High-speed spinning Bioreactor Think About It PMPs are expensive to develop. Chloroplasts have their own DNA and their own protein-making machinery. Isolating the Protein Once the plant has grown and made lots of the protein we want. From there. Some researchers are investigating the use of chloroplasts for making PMPs. Extracting and purifying the protein are often the most expensive steps in making a PMP. Molecules (ligands) attached to the beads adsorb the desired protein. Cultivating plants as protein factories is a good example of how biotechnology uses living organisms to serve humanity. companies plan the entire manufacturing process from start to finish. leaves. The ligand holds the desired protein in place while all the undesired materials in the protein mix are washed off the column with a change in pH or salt concentration (it is the chemical or ionic change that causes materials to leave the column). we might then grind the extract from the oil body. If it’s in a leaf. This would isolate the introduced DNA within the chloroplast—providing a measure of confinement for the PMP. Chemical extraction: Solvents attract the proteins and exclude the cell’s other molecules to leave a crude protein mix.

a healer might have used leaves. Centuries ago. © Kam Yu / Masterfile Genes and Proteins Every living cell contains genes. Insulin lets glucose in the blood enter cells where it can be used for energy. Aspirin production begins with a chemical found in willow bark.Why are people interested in PMPs? Worth Effort the H umans have used plants as medicines since the earliest times. Some proteins help to make new cells. plant-based medicines make up a much smaller portion of the modern medical arsenal. if you were sick. or roots to treat your ailment. but a modern twist may change that. Even today. Researchers are studying how monoclonal antibodies can help diabetics. and others prevent disease. Using plants in this way offers several benefits. Many human diseases arise when the body does not The pancreas secretes a small protein called insulin. Today. some break down food for digestion. Scientists are using plants to produce a wide range of high-tech medicinal proteins. Genes hold the information for cells to make proteins. while morphine comes directly from a particular poppy. about half of our medicines come from chemicals extracted from plants or the plants themselves. bark. Until about 1900 or so. 6 Planting for the Future: Plant-Made Pharmaceuticals . many doctors kept gardens of herb plants that they used as medicines. which the cells use for a variety of purposes.

. using cultures of genetically engineered bacteria. As a result. People with Type 1 diabetes must take daily insulin injections. fertilizer. To use them. pharmaceutical companies should be able to meet the growing demand for biotech drugs. a few seeds from genetically engineered corn or wheat plants may be grown into hundreds of thousands of identical plants in less than two years. Unfortunately. In addition. However. When traditional biotechnology companies need to boost production. they must find a way to make it in existing factories or build a new facility. Producing therapeutic proteins in plants could change the way certain drugs are made. yeast. but manufacturers must still Did You Know. By contrast. making medicines is not something that just anybody can do in his or her spare time. much more. Protein made from animal cell cultures for use in medicine are often contaminated with viruses. Most of these viruses are harmless. this one will take time. HVC. spend a lot of time and money Cultures of Chinese to remove them. and other substances that might harm people who take the PMPs. Sometimes biotech firms rent manufacturing space from other companies or hire another organization to make their product for them. Most protein medicines today are made in factories or laboratories. the body does not make enough of a small protein called insulin. and about 20 universities. handling. HBV) • No human pathogens Antibody • Expensive • Production capacity limited • Cost reduction • Large-scale production Enbrel • Capacity shortage • Large-scale production Factor VIII • Worldwide shortage (40% of hemophilia patients have access to the product) • Large-scale production: accessible to all patients Insulin • Too expensive for non-industrialized countries • Cost reduction: accessible to all non-industrialized countries if they need less! Using plants. protein drugs are not cheap. . it’s farming.000 per gram! (A gram is the mass of a standard paper clip. Only about half a dozen companies worldwide. Only companies with experience making medicines are permitted to manufacture drugs of any kind. or who know farmers. the potential benefits—safer. biotechnology companies must buy sophisticated equipment or construct special buildings. These genes instruct the cell to make a desired protein. For example. Soil. Contamination hamster ovary cells by viruses is not a problem produce a wide variety when plants are put to work of human proteins.Future Farmers of America? Plants: a solution to the shortfall in manufacturing capacity for new drugs Molecule Limitations Plant Solutions Hemoglobin • Blood demand increases. or animal cells. Yes. Most agriculturally useful plants are inexpensive. —Angelo DePalma Your World 7 Canadian Food Inspection Agency make proteins correctly.) One of the goals of biotechnology is to make medically useful proteins (also called therapeutic proteins) in plants at a lesser cost than traditional mammalian cell culture techniques. Cell cultures are extremely expensive to operate. making protein medicines. Companies must prove their protein-making plants are free of disease. or plant less Some of you who live on farms. growing. but it’s much. chemicals. • Large-scale production Donors decrease • Serotypes incompatibility • No concern about serotypes • Security concerns (HIV. and hire highly trained scientists and engineers to operate everything properly. Producing proteins in plants is quite a new idea. All these solutions take time and are very expensive. may get excited about the prospect of producing valuable proteins in your backyards. Plants turn out to be ideal for making biotech medicines. and harvesting must conform to standards that are much stricter than for food plants. in one form of diabetes. companies producing medicines in green plants need only sow more if they need more product. Some cost as much as $50. these plants won’t be sold on the market the way current crops for food and animal feed are. Like all new ideas. easy to grow. more accessible medicines—are worth the effort. and simple to produce in large quantity. Scientists give these cells new genes—genes that are “foreign” to the cells.. Thanks to greenhouses and advanced agriculture. water. Organisms containing foreign genes are known as transgenic organisms. are working on it.

corn. such as alfalfa. a breaksite. new crop.S. At zoos—and many other places—confinement is the key to people feeling secure. growers or permit-holders must surround their crops with a 50-foot buffer zone of fallow (unplanted) land. Unfortunately. APHIS see if they really can harvest the proteins they want. Department of Agriculture (USDA). explain pharmaceutical crops. The all the potential benefits of PMPs will go to waste if companies can’t assure the governcompany must describe the ment and the public that the plants are safe. the government issued permits inspectors also must visit each for just 34 research plots that covered a total PMP production site at least of 130 acres. Companies are concontained. and before planting strategies? soybeans. the industry is still in exactly how the crop will be the experimental stage. use separate equipment. In accordance with regulatory requirements. to produce proteins to at the grower’s be used for medicines. five times throughout the Every step of the PMP process—from hangrowing season and twice dling seeds to planting and harvesting plants more in the subsequent year. It’s easy enough to keep a snake in PMP compaThink About It a cage. list all new genes. In 2002. the ability to 8 Planting for the Future: Plant-Made Pharmaceuticals . but you might not want a harmless garden snake slithering around your ankles either.Is it safe to produce PMPs? Down on th Uncle Sam Is Watching W ould you visit a reptile zoo that didn’t have any cages? The spitting cobra exhibit alone should be enough to keep you home. While many people see a big future for and. to extracting proteins—is already carefully regulated by some combination of the Food and Drug Administration (FDA) and the U. most importantly. As you learn in other artito obtain a perthe potential for human cles. Some plants for PMPs are being grown only in greenhouses. but other things are harder nies are required How can the government to control—such as fields of and companies control plants.S. ducting research with different plants just to As a final precaution. Department of Agriculture that is dedicated to protecting crops from pests and diseases. tion process is lengthy. scientists are now using mit from APHIS error in confinement plants. Currently. the arm of the U. like tractors. but if all plants grown for PMPs are raised this way. in order to first work with a PMP company. growers are required to complete a training course given by the Animal and Plant Health Inspection Service (APHIS). and separate storage facilities for PMPs. The applicathrough that could save lives.

—Chris Woolston Three Models For PMP Production Most companies that produce PMPs set up their operation in one of three ways: • Hire a firm to develop and grow genetically modified plants that produce the protein they’re interested in.000 bushels of soybean crop were in the warehouse. ProdiGene had to buy up and burn 155 acres of surrounding corn that might have been pollinated by its plants. But these areas are popular with growers for a reason. Industry and the government are learning from experience how to fine-tune their procedures. By the time anyone suspected a problem.000 bushels of soybean in that warehouse were destroyed. Two incidents highlighted how diligent everyone involved needs to be. the farmer oversees everything from planting to delivering the harvested crop. • Develop the modified plant themselves and then hire a farmer to sow it on his land. Under orders from the USDA. In that case. A cornfield in Alaska would not be very productive. Will America’s Breadbasket ever become a medicine factory for the world? The science is there. ProdiGene. a Texas-based company. daisies. the entire 500. the 500. All the people involved—from government officials to leaders of biotechnology companies to workers on the ground—are still trying to find the best approach. it will be too late to crossbreed with other crops. • Buy land and set up an operation to do absolutely everything. A few volunteer corn stalks (produced from corn seeds from the previous crop that should have been weeded out by the farmers) sprouted up with the soybeans. Other measures that growers take to ensure the safety of the food supply include planting pharmaceutical crops several weeks after other nearby crops start growing. The technology is still in its infancy.he ‘Pharm’ expand production easily—a main benefit of PMPs—will be limited. everyone on the farm is a company employee. scientists know far more about the genes of cultivated crops such as tomatoes and corn than of. By the time these plants start producing any pollen. used fields in Nebraska and Iowa to grow its plants. Other people say this type of research should be kept out of agricultural hotbeds such as Iowa or Nebraska. Some people believe PMP companies should plant only nonfood crops. Your World 9 . In Nebraska. and ProdiGene had to pay the bill along with a hefty fine. say. A similar problem cropped up at a test plot in Iowa. soybeans were grown in the same field where pharmaceutical corn had been grown the previous year. However.

Which plants are used for PMPs? the Chosen Ones I n the world of plant-made pharmaceuticals. complex proteins used to treat a variety of chronic conditions and life-threatening illnesses. Geography is also sometimes a factor—for example. and bananas are some of the crops being studied. as pure. so it’s not surprising that the search is on for a better alternative. Second.200 per gram of MAB produced in cell culture. confinement can be accomplished by removing the male structure before it begins to shed pollen (detasseling). as well as in several European and Asian countries. About two-thirds of PMP research involves using corn. Other crops have been studied for MAB production. lupine. But how do scientists decide which plants to use and how and where to grow them? Plant-made pharmaceutical (PMP) research is under way in the United States and Canada. are typically produced in cell culture. including large quantities of MABs. the large size of corn pollen allows it to settle quickly. potatoes. rice. sugarcane. lettuce. and research on other crops continues today. say. Finally. corn can also be bred to be sterile. few wild grass species in the United States can crosspollinate with corn. Third. Corn. carrots. Some people worry that plants containing therapeutic proteins might become mixed with plants used for food and feed. tobacco. fold. Monoclonal antibodies (MABs). which is a practice currently used by PMP companies. Researchers point out that we know a great deal more about crop plants than about other plants—including how to confine them. and work as well as one produced in culture. alfalfa. Although corn has a perceived disadvantage—pollen grains containing genetic material could become wind-borne to neighboring fields—it has unique confinement advantages. how do researchers choose the plants they’ll work with? For example. and researchers decide which plant to use based more on economics than on matching plants to specific diseases. a lilac bush? Currently. moss. but so far none has been identified as a suitable host. spinach. Let’s consider an example. Extracting the protein from the plant must be doable—physically possible—in a cost-effective way. barley. time-consuming. And the cost is favorable—$80 to $250 per gram of MAB produced in corn. why would a scientist choose the alfalfa plant instead of. and store large proteins. but they can also be produced in corn. compared with $350 to $1. and expensive process. Excessive water content makes potatoes a poor candidate for . corn has separate male and female flower structures. in the event that corn is not sterile or detasseled. Researchers also consider whether the final processed protein will be as safe. First. researchers might pinpoint a location where a disease is prevalent and then find a crop that is common in that area. tomatoes. manufacturing therapeutic proteins is a challenging. There is no one perfect plant in which to produce every therapeutic protein. safflower. Selected plants of course must be able to express 10 Planting for the Future: Plant-Made Pharmaceuticals and store the desired protein until it is ready to be processed without it breaking down or being degraded (and thus becoming less effective). Finally. which inhibits it from traveling great distances to cross-pollinate. soybeans. duckweed. Corn can efficiently assemble. every plant species has unique advantages and disadvantages that researchers weigh in deciding which to use.

The DNA section that precedes the gene is called the “promoter. The Right Place Remember. Pharmaceutical companies know that an average of 1 gram of monoclonal antibody is produced annually per kilogram of corn seed: 1 g MAB 1 kg corn seed They also know that an average of 175 bushels of corn can be produced per acre and that there are 25. —Kathleen Wildasin From Field to Pharmaceutical You’ve been asked to grow monoclonal antibodies (MABs) in corn and to estimate how many seeds and how much land will be required to grow the seeds before beginning the project. scientists can target where they want the protein to accumulate.7 kg MAB acre Now it’s your turn to do the math. where the plant stores the protein. for instance.000 g ~2. the scale of production. the “termination sequence.445 g MAB x acre 0. storage. He estimates that 150.7 kg MAB = ~167 acres —KW Your World 11 . the gene for the desired protein.000 g = 450 kg (b) How many acres of land must be planted in order to achieve this level of production? 450 kg x 1 acre 2.” It is like an “ON/OFF” switch because it determines when and where the protein is made in the plant. Leaves in tobacco and alfalfa plants and tubers in potatoes have also been sites of PMP production. and a wide variety of equipment (such as grinders.445 kg seed acre Knowing this information. Your project director has given you a few clues about what is already known about MAB production.445 g MAB acre But only an average of 60 percent of the MAB can actually be recovered and purified: 4. and whether drugs or plant-made vaccines are being produced. can be inserted into the plant in any random location. it’s easy to figure out how many grams of MAB can be produced per acre: 4. and the addi- tional steps required for protein extraction and purification in soybeans makes them less attractive than corn.667 g MAB acre x 1 kg = 1. The DNA section that follows the gene. whereas other promoters are “ON” only when they are in specific tissues.4 kg of seed per bushel.000 patients? 150. flanked by DNA sections that regulate gene activity.445 kg corn seed acre x 1 g MAB 1 kg corn seed = 4. the protein is usually mechanically extracted from the plant. but it’s your job to do the final math.MAB production.000 patients x 3g patient x 1 kg 1. Your project director tells you that 3 g of purified MAB are needed to treat one patient for one year. (a) How many kg of purified MAB must be produced each year to meet the needs of 150.000 patients will need treatment each year with your purified MAB.” signals the end of the gene sequence. In PMP production. blenders. Let’s get started. the protein is sometimes kept inside of the plant cell and processed along with the plant material into a powdered form. Matching the right protein to the right plant is a crucial decision for the success of plant-made pharmaceuticals. Some promoters are “ON” all of the time in all tissues. the protein’s ability to withstand the processing procedure so that the desired quantity can be harvested.4 kg seed bushel of corn = 4.60 = 2. and transport. Seeds are most often targeted for PMP production because they are good vessels for protein accumulation. In plant-made vaccine production. and large-scale separating systems) is used. scientists may decide to place the powder into gelatin capsules for oral administration. Later in production. The following calculation shows how much corn seed can be produced on each acre: 175 bushels of corn acre x 25. Picking the Process The method of protein processing depends on the plant being used. By choosing a promoter that is specific to a particular location.

a sophomore in Arlington. Like Meristem. it leaves Troy vulnerable to any pathogens—such as diseasecausing viruses or bacteria—the pig carried. And by making gastric lipase. But the Texas firm ProdiGene has already started marketing a corn-grown enzyme used 12 Planting for the Future: Plant-Made Pharmaceuticals .000 children and young adults worldwide who have cystic fibrosis may soon have a better option. To avoid malnutrition. a genetic disease that makes his body produce thick mucus that clogs his lungs. If a French company called Meristem Therapeutics is successful. Troy has a mild case of CF. more effective treatment for cystic fibrosis patients. Virginia. Troy and the other 70. Because his medication is derived from pig pancreases. By using genetically engineered corn to develop a protein called lipase. That mucus also obstructs his pancreas. the company produces a medication sturdy enough to keep working even after a trip through the stomach. the company bypasses the potential problems of animal tissue. has cystic fibrosis. because pancreatic proteins take a beating in the highly acidic human stomach. You can’t buy PMPs at your local drugstore yet. For some patients. some are creating PMPs to treat diseases.T roy Mashburn. Others are using plants to produce drugs or vaccines to prevent diseases from happening in the first place. he has to take as many as 20 pills a day. Currently in European clinical trials. keeping crucial enzymes from reaching his intestines to help digest his food. But that therapy isn’t perfect. He leads a pretty normal life and even plays on the school’s JV ice hockey team. the medication doesn’t work at all. And the medicine’s effectiveness doesn’t last long. Meristem is just one of the many companies hoping to harness plant power in the fight against disease. Troy uses enzyme replacement therapy. But like other CF patients. the product could mean safer.

” a class of drugs that In fact. and injects these back into the patient to trigger an immune response against malignant cells. Or cholera.3 fucose and xylose includes such proteins as enzymes. California’s Large Scale Biology Corporation is testing individualized vaccines that train the immune system to fight cancer— in this case. They already know how to do this for some plant-made pharmaceutical compounds planned for human use. the product eliminated bacteria to take advantage of plants’ superproductivity for up to a year. hepatitis B.200 live tis.” them an especially attractive traditional vaccines whose proThese 12 million people option where you need lots of teins have been manufactured in special protein to treat the disease. researchers are focusing on the effect the two additional plant N-glycan carbohydrates may have on medicines used in people. Clay specific proteins. and some plant-made pharmaceuticals are in the very early stages of testing in people. or deactivate. The extra carbohydrates in plant N-glycans may cause an allergic reaction in people. researchers are learning how to turn off. To create safe and effective pharmaceuticals in plants. Already in the they are carriers. is trying early trials. feasible to produce just tiny amounts of very —Rebecca A. and antibodies. The that stimulates fat burning. by creating a gel that people could spread on Another of the company’s tobacco-grown their skin to reduce the severity and duration products—this one in the form of nose of genital herpes symptoms. at non-vaccine preventatives. (A different form drops—could prevent the common cold. In Epicyte Pharmaceutical. their immune system to do the About one in every 20 The fact that PMPs could be protection. In the creation of plant-made pharmaceuticals. Proteins that aren’t folded correctly don’t work the way they are expected to. purifies it. Laboratory testing has indicated that these compounds should be safe in humans. non-Hodgkin’s lymphoma—that is already present. The need large quantities of protein—such as for a product bonds to decay-causing bacteria and topical gel—for treating a disease. plants could Using plants could also make it financially turn out to be people’s best friend. animal Nglycans have two additional substances (α 1. The core of the N-glycan chain is identical in both animals and plants. of every 3. They play an important role in protein folding. Crohn’s disease. Hundreds of studies are under way to investigate PMPs’ ability to be used to tackle everything from life-threatening diseases like cancer and HIV to less serious problems like tooth decay and the common cold.Why Do N-Glycans Matter? to produce insulin for diabetics. The company extracts proteins from a patient’s own cancer cells. the plant N-glycans can carry these extra carbohydrates and pass them along into the newly created proteins contained in the medicines. A Canadian comResearchers are also looking of all Americans). horPreventing Disease mones. However. Researchers are Other researchers are using PMPs to prealready working on plant-made treatments vent disease. are usually unaware that a new way. and creates a product that dental Sometimes it’s individual patients who hygienists and patients apply to teeth. California’s prevents them from sticking to the teeth. where millions of people diarrhea. and Caucasian births (in one need long-term treatment. grows identical proteins in tobacco. They’re abnormal “CF gene. CF That’s the case with chronic pipeline are plant-made vacoccurs in approximately diseases like rheumatoid arthricines for such conditions as one of every 3. When plants are used as factories to create medicines.) In the fight against disease. cancer.6 fucose and xylose sugar residues) not found in plant N-glycans. these carbohydrates. What diseases can PMPs combat? Potent Plants N-glycans are carbohydrate chains found within the proteins of both plants and animals. —Joene Hendry Your World 13 . Such tailor-made vaccines could one day offer an alternative to the all-out attack of one-size-fits-all chemotherapy. And plant N-glycans carry two additional carbohydrates not found in animal N-glycans (α 1. for example. is testing a safflower to economically produce a peptide plant-made product to battle tooth decay. The product company uses tobacco to grow an antibody. Treating Disease PMPs could eventually be used to treat just about any disease currently treated with drugs called “biologics. of this virus causes cold sores. Some are looking at vaccines for a long list of conditions—Alzheimer’s disthat run interference with people’s own antiease. bodies rather than triggering a Did You Know? and many more. These aren’t edible Americans is an unafquicker and cheaper to manuvaccines—fruit or other food fected carrier of an facture in large quantities makes with vaccines built in. pany called SemBioSys Genetics A California company called is using genetically engineered Planet Biotechnology. could one day become medicine for people who are dangerously overweight.900 live births take obesity. sugar residues). for example.

Soon Stomp had found a way to genetically engineer the fast-growing weed to produce therapeutic proteins.” she says. Clay . don’t try to be an orchid.” confesses Stomp. Once she felt the company was on solid ground. Inc.D. developed a process that could produce safer. I knew the Latin name of a flower but not that it was a daisy. Undaunted. “Doing biotechnology on trees is like doing biomedical research with whales. “I always come back to plants!” —Rebecca A. Stomp simply created one. a tiny aquatic plant that grows in swamps. she decided that tree-based biotechnology was an area with lots of potential but not lots of competition. Since the university holds the patent and investors own the company. The only problem? There was no duckweed industry to support her research. it may not. she spends her days learning about plants and sharing her discoveries. not what you think you should. then a master’s degree in biochemistry and biophysics from Connecticut in 1981.Career Profile A ccording to Anne-Marie Stomp. Raised in small-town Connecticut by parents obsessed with gardening. And that’s just what students should do.” She pauses. Stomp urges. “Find a desert and be the best cactus you can be. Thanks to that business-like attitude. “As a kid. “If you’re a cactus. I would go on walks with my mom and she would identify plants by their Latin names. she’s doing what she loves. she put her entrepreneurial spirit to work again by launching a biotech company called Biolex. and created a popular seminar called “Anatomy of a Start-up Company” to share with students what she learned along the way.. Developing what she calls a “market entry strategy” for herself. Do what you want. An entrepreneurial approach helped Stomp narrow her focus once she hit graduate school. Then she discovered why so few people were in her field. Stomp loved not only plants but also animals. The following year. Growing up. investigating duckweed’s interactions with other swamp-dwellers. Just as she did as a child. to commercialize her discovery. explaining that trees are too big and slow-growing to make 14 Planting for the Future: Plant-Made Pharmaceuticals AnneMarie Stomp. she convinced timber companies to pay for her graduate work. and finally a doctorate in botany from North Carolina State in 1985. Now Stomp’s back to basic science. In 1998. being a botanist is in her genes. she took a leave of absence and spent the next three years doing everything from designing the company’s labs to raising money to acting as interim CEO. Her training reflects those diverse scientific interests: She earned an undergraduate degree in food science from the University of Connecticut in 1973. Stomp explains. The solution was duckweed. North Carolina State University good research subjects. rocks. Stomp returned to academia. That research may lead somewhere. and every other aspect of nature. cheaper forms of life-saving drugs. “See?” she asks. In the meantime.” Today Stomp is not only a hard-core gardener herself but also an associate professor of forestry at North Carolina State University in Raleigh. Associate Professor of Forestry. she won’t get rich. She does have the satisfaction of having created 40 jobs. Ph.

taxonomic identification). Describe a positive and negative microbial bioassay result. Today. decant sample into labeled Epitube. negative. Make a 25 percent weight by volume plant/solvent mixture. indicates the presence of an antimicrobial substance in the disk saturated by the test extract. a. does this necessarily mean that the natural product is not active? Explain. what is the next step in the drug-discovery process? Explain. allow to sit for 10 minutes.a. Answer these questions: What is the study of a natural product? What is a bioassay? Why is it necessary to mass out the sample and measure out the volume of alcohol? What is an extraction? What is a voucher? Why is it important to document samples in this manner (i. label. with support for development and training from SCCBEP and BABEC. Allow culture to soak into agar surface for at least 10 minutes. Objectives (Record this in your research notebook. A clear area of no growth around the disk. the drug-discovery process involves a sophisticated array of biological assays.” which range from live animal tests to cell culture methods to enzyme assays. and place overnight in a dark environment.. voucher specimen)? Why is it necessary to use a positive and a negative control when performing a microbial assay? What are the expected results of the respective controls (i. and store. Inoculate a cell culture (bacteria or yeast) in a sterile culture tube. If your compound shows a zone of inhibition (halo). Put boat into an incubator to dry. Photo from comstock. One of the simplest assays for antimicrobial activity is the spot disk assay.Activity Microbial Bioassay A number of approaches have been historically used to find leads for new and potentially useful biologically active natural products. Introduce sodium sulfate (desiccant). such as by chromatographic means. Use sterile forceps. Carry out a research project on unknown plant specimens. Your World 15 .e. Procedure Day 2 – Extraction drying and cell inoculate Decant extract (solvent) into clean beaker. then applied again as the mixture is purified. These assays are typically first used to identify a bioactive crude extract. remove. spread to create an even distribution (lawn).) Describe the steps involved in the isolation of a natural product. Spot disk array (area of halo) Procedure Day 3 – Read bioassay plate Read results and record a drawing of what the agar plate looks like in your research notebook. date.com. Silver Creek High School. and extract disks onto the agar plate. Be able to carry out a microbial bioassay. place positive. An aliquot of a test solution is applied to a filter paper disk. and place in aluminum boat (label boat).e. then placed on an agar plate that has been preinoculated with a test microbe. Qualitative results Quantitative results Negative control Positive control Unknown Activity © Mark Okuda. dip an assay disk into extract. cover with aluminum foil. Understand the use of positive and negative controls. to correlate the activity with one particular substance. Describe and practice sterile microbial techniques.. or the zone of inhibition. or “bioassays. and place in a 37°C incubator. measure the distance from the rim of the extract disk to the outer margin of the halo. This “bioassayguided fractionation” yields a pure sample of a molecule that an organic chemist analyzes for structure and that pharmacologists investigate for use in a medicine. Using sterile forceps.k. Cover with aluminum foil. introduce 50ml of a 24-hour-old cell culture onto agar plate. Using sterile technique. Use the table below. If a halo is present. Procedure Day 1 – Extraction and cell culture Make a voucher sample of your specimen (note location. halos)? What does it mean if a halo appears around an experimental assay disk? If a halo does not appear around the disk. in terms of zones of inhibition.

Maize: Indian corn.org/> Growing genetically engineered plants for this purpose is sometimes called “molecular farming” or “pharming.bio. Pfizer Foundation Pfizer Inc Physiome Sciences.ucla.com/businessbriefing/pdf/lifescience2002/publication/daniell.htm#PharmFarming> “Production of Antibodies. Time. Biopharmaceuticals. Express: (v.Monsanto.html> dient believed to provide health benefits. Schering-Plough Research Institute State of Oklahoma Syngenta Biotechnology. Inc. Department of Commerce U.aenews. Biomass: Plant materials and animal waste used especially as a source of fuel. Inc.edu/July02AENews/July02AENews. Massachusetts Biotechnology Council MdBio Merck & Co. protein-based drugs and vaccines. but subscription is free. Inc. Ceres. Inc. <http://www. Inc. Agriculture.org/pmp/> message into a molecular product.) To translate a gene’s Biotechnology Industry Organization.edu/biol/bio1int. Inc.htm#protein> <http://www.wsu.edu/ls3/tutorials> prescribed by a doctor for proper doses. University of Ulster U. Inc.nhmccd. Johnson & Johnson Edward Lanphier Ligand Pharmaceuticals.org/teen/diseases_conditions/digestive/cystic_fibrosis.” Therapeutic protein: A protein that is medically useful. Ernst & Young LLP Genentech. Connetics Corporation CV Therapeutics. Monsanto Fund National Institutes of Health National Science Foundation Nektar Therapeutics Northwestern University’s Kellogg Center for Biotechnology Oak Ridge National Laboratory Onyx Pharmaceuticals.php3?storyID=3D68 (Have to subscribe. Inc.lsic. “Pharm Farming: It’s Not Your Father’s Agriculture. Wiggans Wyeth . Inc. Inc. Inc. Inc. Fact Sheet.” by Henry Daniel <http://www.” by Allan S. Centocor.org/buzz/display.. and Edible Vaccines in Plants. May 26. we say “corn” instead of “maize. and the Environment Biologics: Drugs and vaccines developed with the use of living organisms. Inc.” Nutraceutical: Any food or food ingre- Cystic Fibrosis for Teens (The Nemours Foundation) <http://kidshealth. 2003.usda.aphis.pdf> USDA and Biotechnology: Questions and Answers <http://cofcs66. Aventis American Society for Microbiologists Biogen. Biotechnology Industry Organization Burrill & Company Cell Genesys. Movies & Interactive Tutorial Links <http://science. Pew Initiative on Food and Biotechnology <http://pewagbiotech. Plant-made pharmaceutical (PMP): A Monsanto Web Site <http://www. Alkermes. Genzyme Corporation Government of Canada Inspire Pharmaceuticals. Felsot <http://www. Department of Energy Utah State University Thomas G.com> medically useful protein that is extracted from a plant genetically engineered to express that protein.html> The Biotechnology Institute would also like to thank its 2002 Donors and Campaign Contributors.S. In the United “Cures on the Cob. Ortho Biotech.) Biology/Animations. Inc. Inc. Inc.S. Edible vaccines: Vaccines produced in food crops that can be eaten as Resources •• •• • Glossary Harvest More Info AgBiotech Buzz.gov:80/biotech/Bio_qa. Amersham Biosciences Corp Amgen.wmrc. Inc. States. Sangamo BioSciences. InterMune Pharmaceuticals.You’ll find— • Teacher’s guide • Activity supplement: student and teacher procedures • Links • Information on subscriptions and previous issues • Downloadable teacher’s guides for previous issues These issues of Your World are available to download FREE— • Exploring the Human Genome • Gene Therapy • Environmental Biotechnology • Industrial Biotechnology • Plant Biotechnology • Health Care.” by Margot Roosevelt. Inc. Roundtable: Is There a Pharm in the Future? <http://pewagbiotech. Abgenix.