Humulin is synthetic human insulin prepared by using Recombinant DNA Technology. It is one of the best example of Genetically Modified Microorganisms.

1. 1
GMO: Humulin
History, Methods And Risks
MS. GAURAVI RAUT
M.SC. BIOTECHNOLOGY
PATKAR-VARDE COLLEGE

2. Introduction
Humulin
History
Structure of human insulin
How do they make insulin from recombinant DNA
Types of Humulin
Risks
Advantages of Humulin Over Insulin
Reference
INDEX 2

3. Genetically modified microorganisms: Alteration of an organism’s gene for practical purpose
Performed by modifying an organism’s own DNA or introducing new DNA to perform desired
functions
GMOs have emerged as one of the mainstays of biomedical research
Recombinant DNA technology: uses genetic engineering to introduce genes into
microorganisms and other cells and help to produce metabolites of commercial or medical
importants such as insulin, vitamin, amino acids or enzymes
Most reason to prefer microorganisms:
 Grow rapidly
 Easy to cultivate
 Environmentally friendly than conventional chemical synthetic method
Pharmaceutical products:
 Recombinant hepatitis B vaccine (GM Baker’s Yeast)
 Injectable insulin (GM Escherichia coli)
3INTRODUCTION

4. Synthetic human insulin used for treating diabetes
Recombinant DNA technology
Prior to its development, diabetics used insulin isolated from pig and cow pancreases
Recombinant pharmaceuticals
4HUMULIN
Inserting genes
from one species
into host species
(yeast or bacteria)
The genes code for
a desired product
Therefore
genetically modified
host organisms can
be grown and used
as a kind of living
factory to produce
the product
In this case, genes
coding for human
insulin are inserted
into bacteria
Bacteria produce
insulin which is
harvested and used
as the active
ingredient in
Humulin

5. In 1982, Lilly's Humulin: first genetically engineered drug approved by the US FDA
FDA approval: five months after application was made by Eli Lilly and Company of
Indianapolis, Ind.
Normally, such approval takes 20 to 30 months
First drug produced by genetic engineering techniques to gain the FDA approval for
human use
Efforts: four years ago, after scientists at the City of Hope National Medical Center in
Duarte, Calif., and Genentech Inc. in South San Francisco first succeeded in producing
it in the laboratory
Inserting human genes responsible for insulin production into E. Coli bacteria, thus
stimulating the bacteria to synthesize insulin
Humulin had proved to be safe and effective in FDA clinical trials involving more than
400 patients
Today, available in both a vial and a disposable prefilled pen: helps in choosing a
convenient way of taking humulin
HISTORY 5

6. Hormone produced by β-cells of islets of Langerhans of pancreas
Peptide hormone produced by pancreas and is a central regulator of carbohydrates and fat metabolism in the body
It controls blood sugar lever and when there is less secretion of insulin, it results in diabetes (high blood sugar level)
Insulin consists of a 21-amino acid A chain and a 30-amino acid B chain, linked by two disulfide bonds. It can be
produced either by generating the chains separately and chemically combining them or by creating a single-chain
precursor, human proinsulin and cleaving out a 35-amino acid connecting peptide
 For manufacturing the drug, the proinsulin route is favored because it requires a single fermentation and isolation step
6STRUCTURE OF HUMAN INSULIN

7. At first, suitable vector (plasmid) is isolated from Escherichia coli and then it is cut open by restriction endonuclease enzyme
The gene of interest (i.e. insulin coding gene) is isolated from β-cells and inserted in open plasmid
Plasmid and gene of interest are recombined together by DNA ligase enzyme
This recombined plasmid is inserted into suitable host cell (i.e. Escherichia coli) and now this recombined host cell
starts producing insulin hormone
Then extraction of recombinant gene product from Escherichia coli is done
Purification of humulin
7HOW DO THEY MAKE INSULIN FROM RECOMBINANT DNA?

8. Basic Steps Involved In Humulin Production

9. 9
These genes are inserted by the side of β-galactosidase gene of the plasmid
The recombinant plasmid were then separately transformed into Escherichia coli host
The recombinant host produced proinsulin chains i.e. fused β-galactosidase A chain and β-galactosidase B
chain separately
The proinsulin chains A and B were separated from β-galactosidase by treatment with cyanogen bromide.
The detachment of proinsulin chains from β-galactosidase is possible because an extra codon from
methionine was added at N-terminal of each gene for A and B chain
After detachment, A and B chains are joined invitro to reconstitute the naive insulin by sulphonating the
peptide chains with sodium disulphonate and sodium sulphite
Hakura et al(1977) chemically synthesize DNA sequence of insulin for two chains A and B and separately
inserted into two PBR322 plasmid vector

11. 11WHAT TYPES OF HUMULINS ARE AVAILABLE?
For the treatment of high blood sugar (glucose) levels, 3 types of Humulin can help to manage diabetes
Each type varies in onset, peak time and duration
 Short-acting insulin
 It can cover insulin needs for meals
eaten within 30 minutes
 NPH (Neutral Protamine Hagedorn)
insulin
 Intermediate acting insulin
 Slower to act and lasts longer than
regular human insulin
 Short-and intermediate-acting insulins combined to cover
insulin needs after meals, throughout the day, and at night
 Available in a number of different ratios of mixing
 For example,
a) Humulin M3: mix of 30% short acting & 70%
intermediate acting
b) Humulin M5: 50% of both short & intermediate acting
Humulin R Humulin N Premixed Humulin

12. 12
Hypoglycemia
Hypokalemia
Edema
Lipodystrophy
Weight gain
Immunogenicity
Hypoglycemia
Sweating
Dizziness
Hunger
Anxiety
RISKS

13. Earlier insulin required for diabetes was extracted from pancreas of slaughtered cattle, pigs or salmon.
The process was quite tedious and difficult and yields of insulin would be low.
This extracted insulin in some patients, developed allergy or other side effects due to foreign protein. Due
to disadvantages of animal insulin and advantages of humulin, humulin is regarded superior to animal
insulin.
Humulin is considered better than animal insulin because:
 Absorbed more rapidly and show its effectiveness in short duration
 Causes fewer allergic and autoimmune reactions as compared to animal insulin
 Less expensive than animal insulin
13ADVANTAGES OF HUMULIN OVER INSULIN

14. www.onlinebiologynotes.com
www.biospectrumindia.com
www.humulin.com
14REFERENCE

15. THANK YOU!!!!!