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INSULIN MANUFACTURING PROCESS
2
INSULIN MANUFACTURING PROCESS
INSULIN PRODUCTION
UPSTREAM
1) Obtaining of human insulin gene
2) Insertion of cDNA of both the chains into plasmid
3) Transfection (Recombinant plasmids enter the bacteria in a process known as transfection)
4) Media and equipment preparation
PRODUCTION
- Fermentation section
- Reaction section
- CNBr cleavage
- Sulfitolsis and refolding
- Enzymatic conversion
DOWNSTREAM PROCESS
Purification by various methods
- RP-HPLC
- Chromatography
- Centrifugation
INSULIN
Mass balance
Insulin is a hormone made by the pancreas
that allows your body to use sugar (glucose) from carbohydrates in the food that you eat for energy or to store glucose for future use. Insulin helps keeps your blood sugar level from getting too high (hyperglycemia) or too low (hypoglycemia).
The cells in your body need sugar for energy. However, sugar cannot go into most of your cells directly. After you eat food and your blood sugar level rises, cells in your pancreas (known as beta cells) are signaled to release insulin into your bloodstream. Insulin then attaches to and signals cells to absorb sugar from the bloodstream. Insulin is often described as a “key,” which unlocks the cell to allow sugar to enter the cell and be used for energy.
Energy balance
Proinsulin Isolation
Purification process
Several Process
Homogenizes
Washer
Centrifuge
Homogenizes
Blending Tank
Rotary vacuum
CNBr cleavage
Solubilizatio
Recentrifuge
Diafiltration unit
Refolding
Sulfitolysis
Replacer
Cation exchanger
Chromatography HIC COLUMN
Enzymatic conversion
Diafiltration unit
HUMAN INSULIN
is
produced from Proinsulin
MANUFACTURING PROCESS
Production of pro insulin takes place in E.coli,
which then further isolated from the cell
Step 1
We transformed E. coli cells, These cells are used to produce the
Trp-LE'-MET-proinsulin precursor of insulin, which is retained in the cellular biomass.
In Bioreactor (Fermentation reaction)
Glucose+Ammonia+Air +Broth +Biomass is added to a bioreacter where fermentation takes place and 30g/litre of E.coli is produced of which 20% is the inclusion body of the total dry cell mass.