PM SHRI KENDRIYA VIDHYALAYA

SHIFT- 1, PATTOM,
THIRUVANANTHAPURAM (2023-24)

BIOLOGY INVESTIGATORY PROJECT

Submitted By
Ananthapadmanabhan A S
XII A
CBSE Roll no- 24602252
INVESTIGATORY PROJECT
BIOLOGY

HUMAN-INSULIN
 CERTIFICATE
It is hereby to certify that, the original and genuine
investigation work has been carried out to investigate about the
subject matter and the related data collection and investigation
has been completed solely, sincerely and satisfactorily done by
Ananthapadmanabhan A.S of Class XII A, Kendriya Vidyalaya
Pattom shift I, regarding the project titled, “HUMAN-
INSULIN”

……………………… …………………
Signature of the Guide Principal’s Signature

Submitted for ALL INDIA SENIOR SECONDARY

EXAMINATION held at KENDRIYA VIDYALAYA Pattom
shift I

……………………… ……………………
Internal Examiner External Examiner
 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.
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. In addition to the laboratory, Macleod
provided experimental animals and his student Charles Best,
who worked as a demonstrator. Macleod instructed Banting on
the 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". 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 unravelled
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.
 BIBLIOGRAPHY

 Biomedical Research and Therapy -

Journal for Medical Biotechnology and Medicine
Incorporating Advances in Regenerative Medicine
 http://bmrat.org/index.php/BMRAT
 The National Center for Biotechnology Information (US)
 https://www.ncbi.nlm.nih.gov/
 Wikipedia
 https://www.wikipedia.org/