CENG 109 class 4
Putting biology into a bottle:
the story of recombinant
insulin
Class announcements
Hand in homework #1 NOW
„ Team formation deadline (18/09)
„ Email questions and results to TA Stephen
stepheny@ust.hk
„ Arrangement for library orientation
„
Recombinant insulin starts a new
chapter for biotechnology industry
Human insulin
The first synthesis of
Aspirin in 1899
Penicillin- the first antibiotic
was discovered in 1928
www.rcsb.org
First recombinant product
hit the market in 1982
Outline of this class
1)
2)
3)
The health care problems for diabetic
patients now and then (70’s)
Windows of opportunities for the new
biotechnology
Technical and regulatory challenges for
the development of recombinant insulin
Diabetes is a serious
health and economic problem
„
„
„
Diabetes affects more than 230 million
people worldwide
Diabetes is the leading cause of blindness in
developed countries, a common cause for
amputation and kidney failure, and increases the
chances for developing heart conditions.
The devastating complications of diabetes are
imposing a huge burden on healthcare
services. It is estimated that diabetes accounts
for between 5% and 10% of a nation's health
budget. Information from the web site of International Diabetes Federation
www.idf.org
Insulin :a standard treatment for
diabetes since 1924
1921: First isolation of
insulin from animal
(Univ. of Toronto)
(Macleod and Banting were
awarded the Nobel Prize
in 1923 for the discovery
of insulin. )
„ 1922: Large scale
production/
commercilization
(Eli Lilly; Novo Nordisk)
„ What was the source of
insulin?
„
Human insulin differs from pig insulin
by 1 amino acid; from cattle insulin by
3 amino acids
Commercial opportunities for the
new technology
„
Advances in new technology
In 1958, the exact sequence of amino acids comprising the
insulin molecule, the so-called primary structure, was
determined by British molecular biologist Frederick Sanger.
In 1967, after decades of work, Dorothy Crowfoot Hodgkin
determined the spatial conformation of the molecule, by
means of X-ray diffraction studies. She had been awarded a
Nobel Prize in Chemistry in 1964 for the development of
crystallography.
Boyer and Cohen devised methods for making recombinant
DNA in 1974.
Rosalyn Sussman Yalow received the 1977 Nobel Prize in
Medicine for the development of the radioimmunoassay for
insulin.
(www. Wikipedia.com)
Commercial opportunities for the
new technology
„
Limitations of animal extraction
Potential allergic reactions
Slaughterhouse would not be able to provide enough
supply of animal pancreas
„
Market size of the proposed product
Increased diabetes patients
Insulin is the mainstream treatment
Animal-insulin is already a mature product
Lucrative markets
„
Insulin is a relatively small and simple
protein
Technical Challenges
“The thrill of it was that nobody had every
done it before. The problem with it was
that no one had ever done it before”
Bruce H. Frank, a retired Lilly scientist who
worked on the development of recombinant
human insulin
Making insulin with
recombinant DNA technology
„
Obtaining the insulin gene
1) mRNA isolated
from beta-cells in
pancreas
mRNA
2) cDNA produced by
Reverse transcriptase
mRNA
cDNA
3) mRNA destroyed
cDNA
4)DNA stranded produced
by DNA polymerase
double stranded DNA
Graphics adapted from “Biotechnology: Demystifying the Concepts” by Bourgaize, Jewell, Buiser
Cloning human insulin gene
in bacterial cells
Graphics from “Biotechnology: Demystifying the
Concepts” by Bourgaize, Jewell, Buiser
Procedure for identifying insulin
producing cells
Graphics adapted from “Biotechnology: Demystifying the Concepts” by Bourgaize, Jewell, Buiser
Scaling up the fermentation was
only the first step in manufacturing..
Eli Lilly licensed the recombinant human
insulin from Genentech. Its expertise
in isolating antibiotics from fermentation
was important in scaling the production
of recombinant protein from bacterial
fermentation
Insulin is more than a chain of
amino acids
New techniques for quality control
„
New physiochemical and biological test
were introduced
Confirm
the primary sequence
Confirm the 3-D structure
Answer questions about immunogenicity (is
there any allergic reaction?), efficacy (is it
performing its biological functions?), and
stability (is it still the same protein after
storage?)
Regulatory Challenges
„
„
„
„
FDA had experiences approving insulin products but
none with recombinant protein products
Public caution has imposed further challenges in early
bioprocesses (e.g. the two-chain method, small scale
fermentation)
FDA worked closely with companies and quickly
expanded its expertise in genetic engineering, resulting
in a quick approval process of the somewhat
controversial product at the time
Humulin was launched in 1982, only four years after the
initial demonstration that human insulin could be
expressed in bacteria using rDNA method.
Revenues of insulin products
More than 20 insulin products available,
most are produced by recombinant routes
„ Insulin remains a billion-dollar drug today.
In 2004, insulin sales brought
„
$3.4
billion for Novo
$2.1 billion for Lilly
$1.1 billion for Sanofi-Aventis
Homework #2 (due 21/09)
1)
2)
Aside from Genentech, there are a number of
biotechnology giants and their global market
shares are displayed in the chart below. By
browsing the company web sites, find out
information of a major human health product
from 3 of these companies. What are the
therapeutic targets of these products? What
diseases do they treat?
Discuss major obstacles in developing
recombinant insulin into a commercial product