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/