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

………………….. ……………………

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.