INVESTIGATORY PROJECT

BIOLOGY
HUMAN-INSULIN

Ananthapadmanabhan A S
XII A
PM SHRI Kendriya Vidyalaya pattom shift I
 CERTIFICATE
This is to certify that this Biology Investigatory Project titled
HUMANINSULIN has been successfully completed by
Ananthapadmanabhan A.S in partial fulfilment of the
requirements for the award of the SSCE 2024 PRACTICAL
Examination in biology

Internal examiner External examiner

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Principal

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 AKNOWLEDGEMENT

I would like to express my special thanks of gratitude to my

Biology Mentor Ms Anitha Ramesh as well as our
honourable Principal sir shri R. Giri Sankaran Thampi who
gave me the golden opportunity to do this project on the topic
“HUMAN INSULIN” which also helped me in doing a lot of
research and to learn new things. I would also like to thank my
parents and friends who helped a lot in finalizing the project
within the limited time frame. I am making this project not only
for marks but also to increase my knowledge
 What is insulin?
Insulin is a hormone which acts as a key regulator of blood sugar levels, ensuring that
the body has a steady supply of energy for its various functions. It is produced by the
beta cells of the pancreas, which is an organ located behind the stomach. Insulin is
released in response to elevated blood glucose levels, i.e. after consuming food.

Insulin plays a central role in maintaining blood glucose levels within a narrow and
healthy range.

Dysregulation of insulin production or function can lead to conditions such as diabetes.

In diabetes, the body either does not produce enough insulin (as in Type 1 diabetes) or
is unable to use insulin effectively (as in Type 2 diabetes). This results in elevated blood
sugar levels, which can lead to various health complications if not properly managed.
In these cases, individuals may need insulin injections or other medications to help
regulate their blood sugar.

during the early to mid-20th century. insulin was extracted from animal sources. The
process of extracting insulin from animals was a crucial advancement in diabetes
treatment during the early times

Problems with cattle derived insulin

While animal-derived insulin was a groundbreaking treatment for diabetes, it had
some limitations. There were slight differences between animal insulin and human
insulin, and some individuals experienced allergic reactions or resistance to animal
insulin. Additionally, the supply of animal pancreases was finite and subject to
variations in insulin content.
 Development of DNA technology
The development of recombinant DNA technology in the late 20th century allowed for
the creation of synthetic human insulin This biotechnological breakthrough enabled
the large-scale production of insulin using bacteria to produce human insulin
Synthetic insulin is virtually identical to the insulin produced by the human body,
reducing the risk of adverse reactions and providing a more reliable source of insulin
for individuals with diabetes. Synthetic insulin has since become the standard for
diabetes treatment.
The development of recombinant DNA technology in the late 20th century was a
groundbreaking advancement in the field of biotechnology. Recombinant DNA
technology allowed scientists to manipulate and combine DNA from different
sources, paving the way for numerous applications in medicine, agriculture, and
industry.
One of the significant achievements of this technology was the production of
synthetic human insulin.

Production of synthetic human insulin

The production of synthetic human insulin using recombinant DNA technology in
the 1980s was a groundbreaking achievement in the field of biotechnology. This
marked a significant advancement in diabetes treatment, offering a safer and
more sustainable source of insulin for individuals with diabetes

Isolation of the Human Insulin Gene:

• The first step involved isolating the gene responsible for producing
human insulin. Scientists identified and extracted the specific DNA
sequence that codes for insulin from human cells.

• Messenger RNA is a molecule of RNA that encodes a chemical

"blueprint" for a protein product.

• The isolated gene contains the code of the human DNA for the
production of insulin.

• The plasmid DNA of the bacterial cell is taken out of the cell.
Insertion into a Plasmid Vector:
• The isolated human insulin gene was then inserted into a small,
circular DNA molecule called a plasmid vector. Plasmids are often
used as carriers to introduce foreign genes into host organisms

• The plasmid DNA of the bacteria is cut out producing plasmid ring
which is an empty segment of the DNA.

• A Restriction Enzyme is an enzyme that cuts DNA at specific

recognition nucleotide sequences known as restriction sites.

Introduction into Host Organisms:

• The recombinant plasmid, now containing the human insulin gene,
was introduced into host organisms, commonly bacteria (Escherichia
coli) or yeast cells. These host organisms would serve as living
factories for the production of insulin.
Expression of the Insulin Gene :
• Once inside the host cells, the insulin gene in the plasmid directed
the host cells to produce human insulin. The cellular machinery of
the host organism, including transcription and translation processes,
was utilized to generate the insulin protein.

Harvesting and Purification:

• The insulin produced by the host cells was harvested and then
subjected to purification processes. These processes were
designed to separate insulin from other cellular components and
contaminants, resulting in a highly purified form of synthetic
human insulin.
Production of Humulin
The cells need nutrients in order to grow, divide, and live. While they live, the
bacterial cell processes turn on the gene for human insulin and the insulin is
produced in the cell. When the bacterial cells reproduce by dividing, the human
insulin gene is also reproduced in the newly created cells.

Discovery of human insulin

Human insulin was discovered through collaborative efforts, and the
credit for its discovery is often attributed to a team of researchers led
by Sir Frederick Banting, Charles Best, James Collip, and John Macleod.
The discovery took place in the early 1920s

1921: Frederick Banting and Charles Best

Frederick Banting, a Canadian surgeon, and Charles Best, a medical
student, conducted experiments at the University of Toronto. Banting had
the idea that if the pancreas could be isolated from its digestive
secretions, the extract might contain a substance that could treat
diabetes. In the summer of 1921, Banting and Best began their
experiments.
1921 (Isolation of Pancreatic Extract):
Banting and best, with the assistance of laboratory technician
James Collip, successfully isolated a pancreatic extract that, when
injected into diabetic dogs, effectively lowered their blood sugar
levels. This extract contained what we now know as insulin.

1922: First Successful Use in Humans

In 1922, the first successful use of insulin in a human patient
occurred. Leonard Thompson, a 14-year-old boy with diabetes,
received an injection of the newly discovered insulin. The treatment
was successful, and the boy's health improved dramatically.

1923: Nobel Prize in Physiology or Medicine

In 1923, Frederick Banting and John Macleod (who provided
laboratory facilities and guidance) were awarded the Nobel Prize in
Physiology or Medicine for the discovery of insulin. Banting shared
the prize money with Charles Best, and Macleod shared it with
James Collip.

.
Masterminds behind the discovery
Frederick banting (1891–1941)

He was a Canadian medical scientist, physician, painter, and Nobel

laureate noted as the co-discoverer of insulin and its therapeutic potential. born
on November 14, 1891
Banting was appointed Senior Demonstrator in Medicine at the University of
Toronto in 1922. Next year he was elected to the new Banting and Best Chair of
Medical Research, endowed by the Legislature of the Province of Ontario. He also
served as Honorary Consulting Physician to the Toronto General, the Hospital for
Sick Children, and the Toronto Western Hospital. At the Banting and Best Institute,
he focused his research on silicosis, cancer, and the mechanisms of drowning.

In 1938, Banting's interest in aviation medicine resulted in his participation with

the Royal Canadian Air Force (RCAF) in research concerning the physiological
problems encountered by pilots operating high-altitude combat aircraft. Banting
headed the RCAF's Number 1 Clinical Investigation Unit (CIU), which was housed in
a secret facility on the grounds of the former Eglinton Hunt Club in Toronto.[15]
John Macleod (1876–1935)

He was a Scottish biochemist and physiologist. He devoted his career to diverse

topics in physiology and biochemistry, but was chiefly interested in carbohydrate
metabolism. He is noted for his role in the discovery and isolation of insulin during
his tenure as a lecturer at the University of Toronto, for which he and Frederick
Banting received the 1923 Nobel prize in Physiology or Medicine.
At the end of 1920, Macleod was approached by Frederick Banting, a young Canadian
physician who had the idea of curing diabetes using an extract from
a pancreas whose functioning had been disrupted. Macleod was not enthusiastic,
because (unlike Banting) he knew about unsuccessful experiments in this direction
by other researchers. He thought it more likely that the nervous system had a
crucial role in regulating blood glucose concentration. Macleod was not initially
impressed by his interview with Banting. However, he came to the conclusion that
it was worth trying because the results may be of "great physiological value," and
granted Banting laboratory space while Macleod himself would be away on
holiday.[11] In addition to the laboratory, Macleod provided experimental animals
and his student Charles Best, who worked as a demonstrator. Macleod instructed
Banting on the accepted method of pancreatectomy to be used on the experimental
subjects.
Charles Best (1899–1978)

He was an American-Canadian medical scientist and one of the co-discoverers

of insulin.
in 1915 he moved to Toronto, Ontario, where he started studying towards a bachelor
of arts degree at University College, University of Toronto. In 1918, he enlisted in the
Canadian Army serving with the 2nd Canadian Tank Battalion. After the war, he
completed his degree in physiology and biochemistry.
As a 22-year-old medical student at the University of Toronto he worked as an
assistant to the surgeon Dr Frederick Banting and contributed to the discovery of
the pancreatic hormone insulin, which led to an effective treatment for diabetes. In
the spring of 1921, Banting travelled to Toronto to visit John Macleod, professor of
physiology at the University of Toronto, and asked Macleod if he could use his
laboratory to isolate pancreatic extracts from dogs. Macleod was initially sceptical,
but eventually agreed before leaving on holiday for the summer. Before leaving for
Scotland, he supplied Banting with ten dogs for experiment and two medical
students, Charles Best and Edward Clark Noble, as lab assistants.
James Collip (1876–1935)

He was a Canadian biochemist who was part of the Toronto group which
isolated insulin. He served as the chair of the department of biochemistry at McGill
University from 1928 to 1941 and dean of medicine at the University of Western
Ontario from 1947 to 1961, where he was a charter member of The Kappa Alpha
Society.
MacLeod was overseeing the work of Frederick Banting and Charles Best in their
search for a treatment for diabetes which they had begun in May 1921. In December,
when Banting and Best were having difficulties in refining the pancreatic extract,
MacLeod freed Collip from his other research to enable him to join the research
team. Collip's task was to prepare insulin in a more pure, usable form than Banting
and Best had been able to achieve to date. In January 1922, after 14-year-
old Leonard Thompson suffered a severe allergic reaction to an injection of insulin,
Collip achieved the goal of preparing a pancreatic extract pure enough for
Thompson to recover and to use in clinical trials. Despite Collip's breakthrough,
Banting was furious as he saw that "Collip's discoveries were not a cause for
celebration but a new threat".[4] At some point between January 17 and 24, Collip
and Banting reportedly had a physical altercation in the labs, supposedly when
"Collip visited Banting and Best in their lab and told them that he wasn’t going to
share the latest extract formulation (which may or may not have had Macleod's
blessing) and that he was contemplating leaving the research team and patenting
the process on his own". A colleague later lampooned this incident with a "cartoon
showing Banting sitting on Collip and titled 'The Discovery of Insulin. Nonetheless,
successful trials were soon completed and the future of insulin was assured.
Banting, Best and Collip subsequently shared the patent for insulin

Conclusion
In conclusion, the exploration of human insulin has unraveled a remarkable journey
from its initial discovery to the contemporary era of biotechnological advancements.
The collaborative efforts of Sir Frederick Banting, Charles Best, James Collip, and
John Macleod in the early 1920s laid the foundation for a groundbreaking treatment
that transformed the lives of individuals grappling with diabetes.
The development of human insulin, particularly the synthetic forms like Humulin,
has significantly enhanced the management of diabetes. The evolution from animal-
derived insulin to the recombinant DNA technology-enabled synthetic insulin
reflects not only scientific ingenuity but also a commitment to improving the safety,
efficacy, and accessibility of diabetes treatment.
Recombinant DNA technology emerged as a pivotal player in the production of
human insulin, allowing for the creation of genetically engineered organisms that
act as insulin factories. This innovation not only addressed the limitations of animal-
derived insulin but also paved the way for the broader applications of genetic
engineering in medicine and biotechnology. Furthermore, the impact of human
insulin extends beyond its therapeutic use. It has become a symbol of the potential
of biotechnological advancements to address complex health challenges. The
intersection of genetics, molecular biology, and medical science has propelled the
development of personalized medicine, gene therapy, and other transformative
approaches. As we reflect on the journey of human insulin, it is evident that the
story is far from over.Ongoing research continues to refine treatment options,
explore new avenues in gene therapy, and deepen our understanding of the intricate
mechanisms governing glucose metabolism. The field of insulin research remains
dynamic, with the potential to unlock further innovations in diabetes care and
related medical domains.
In conclusion, the saga of human insulin exemplifies the power of scientific
collaboration, innovation, and perseverance in shaping the landscape of medical
advancements. It stands as a testament to the ability of humanity to harness the
intricacies of biology for the betterment of lives, and it provides a foundation for
future breakthroughs in the ever-evolving field of medical science.