INSULIN

Introduction to Insulin
Insulinis apeptide hormone, produced bybeta

cellsin thepancreas, and is central to regulating

carbohydrate andfat metabolism in the body.
It causes cells in theskeletal muscles, andfat
tissueto absorbglucosefrom theblood.
Insulin is a small protein. It contains 51 amino acid
arranged in two chains (A and B) joined together by
disulfide linkage.
The molecular origins of insulin go at least as far
back as the simplest unicellulareukaryotes.
Apart from animals, insulin-like proteins are also
known to exist in Fungi and Protista kingdoms.

History of Insulin
Insulin was identified as a substance in

thepancreasin 1869 by German medical student

Paul Langerhans, though its function and properties
were not known.
Its relationship to blood sugar was illuminated by
Polish scientists in 1889, but insulin was no isolated
and specifically studied until the early 1920s.
The first insulin injection given to successfully treat
a diabetic was administered in 1922 in Toronto, and
intense study and manipulation of the protein has
carried on since then.

Human Insulin
Production In Body
Translation and translocation
Folding,oxidation and signal peptide cleavage
Endoplasmic Recticulum export,Golgi transport

and vesicle packaging

Protease cleavage liberate C-peptide
Carboxypeptidase E produces mature insulin

Human insulin production

by genetic engineering

Upstream production of INSULIN

Step 1: Obtaining the insulin gene

Step 2: Insertion of gene into

plasmid
Restriction enzyme and ligase is involve

Step 3: Transfection
Using CaCl and eletroporation treatment

Step 4: Media equipment preparation

Fermentation

- broth containing antibiotic (ampicillin and

lactose)

Downstream production of
INSULIN

Step 6: Isolation of crude products

Cells are removed from tanks and are lysed
Eg.

For enzyme digestion, lysosomal enzyme is used to

digest the outer layer of the bacterial cells and
detergent mixture is subsequently added to separate
the cell wall membrane.

Step 7: Purification of crude

product
Centrifugation is conducted to helps separate

the cell components from theproducts.

Stringent purification of the recombinant
insulin chains must be taken to remove any
impurities. This uses several chromatographic
methods, along with additional steps which
exploit differences in hydrophobicity.

Step 8: Obtaining Insulin Chain

The proteins isolated after lysis consists of the

fusion of -galactosidase and insulin chains

due to the fact that there is no termination or
disruption to the synthesis of these two
proteins as the genes are linked together.

Step 9: Synthesis of active insulin

Two chains (A and B) forms disulfide bonds

using sodium dithionate and sodium sulphite,

and the chains are joint through a reaction
known as reduction-reoxidation under betamercaptoethanol and air oxidation, resulting
in Humulin - synthetic human insulin.

Step 10: PR-HPLC to obtain

highly purified insulin
Reverse-phase high performance liquid

chromatography (PR-HPLC) is performed lastly

to remove almost all the impurities, to
produce highly purified insulin.
The insulin then can be polished and
packaged to be sold in the industries.

THE OPERATIONAL
PROBLEMS IN INDUSTRY

INSULIN AND ITS INVOLVEMENT

IN DIABETES
Insulin is ahormonethat your body

produces to help convert the food you eat

into energy
People with diabetes might need insulin
either because they dont produce enough
insulin or they cant properly use the insulin
that they do produce or both.
Insulin is the hormone that helps glucose
move from the blood into your body cells.

The most common way to get insulin is to

inject it with an insulin pen, syringe, or pump.

Insulin works best when injected into the fat
layer just beneath the skin.Some areas of the
body to use include the:
Abdomen (except a two inch circle around the

belly button)
Thighs (top and outer parts)
Backs of the upper arms

WHAT IS
DIABETES?

Diabetes mellitus, also known as

sugar diabetes, occurs when the
body is either unable to produce
enough insulin or is unable to use
the insulin that is produced.

Types
of
Diabete
s
Type 1 diabetes is a
condition that occurs
when the pancreas
either does not
produce insulin or
produces very little
insulin.

Type 2 diabetes is a
condition that occurs
when the body
develops a
resistance to any
insulin that is
produced or when
the body produces
insufficient amounts
of insulin.

Pre-diabetes is a
condition that
occurs when blood
glucose levels are
elevated but not
high enough to be
considered a
symptom of fullblown diabetes.

How Insulin
Helps Control
Blood
Glucose
Levels

Safety Precautions
Keep unopened insulincontainersin the

refrigerator.
Once the container is open, it can
bestoredatroom temperaturefor as long as the
label permits.
Keep insulin products from becoming toohotor
too cold.
Never use insulin after theexpiration date.
Use needle tips and syringes once and then
dispose of them in asafe containerlabeled
diabetes supplies.

Conclusion
Insulin analogues are just such a product as

they are artificial derivatives of the natural

hormone insulin, designed to improve the
absorption profiles compared to human insulin
after subcutaneous injection.
Of special concern is that the carcinogenic
potential of insulin analogues remains to be
determined on human carcinoma tissue.