Now let us look at a specific example where affinity chromatography is used, namely, in the purification of human leukocyte interferon made in E. coli.
Proteins synthesized in genetically engineered organisms and intended for injection into animals must be stringently purified. Pyrogens from E. coli, including the outer envelope lipopolysaccharide (LPS) must be removed or inactivated. Hence product recovery operations such as affinity chromatography are an important step in the manufacturing process.
Given below is a schematic representation
of the typical steps involved in processing human leukocyte interferon
produced by recombinant DNA techniques. This will give you an idea of where
exactly affinity chromatography is usually involved in the realm of bioprocessing.
HUMAN LEUKOCYTE INTERFERON
|
E. coli EXTRACTION BY MECHANICAL BREAKAGE
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POLYETHELYNEIMINE PRECIPITATION
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AMMONIUM SULFATE PRECIPITATION OF SUPERNATENT
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DIALYSIS OF PELLET
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* IMMUNOADSORBENT COLUMN (MONOCLONAL ANTIBODIES)
|
CATION EXCHANGE CELLULOSE CHROMATOGRAPHY
SO, Thank you everybody, for reading so patiently to what I had to say about affinity chromatography....
I hope you enjoyed the show...
page designed by: Kaumudi M. Bhawe
(Biochemical Engineering Term Project - Fall 1997.)