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US 8,956,812 B2Granted
Process for the preparation and purification of recombinant proteins
AssigneeBharat Biotech International Limited
InventorsKrishna Murthy Ella, Srinivas Kannappa Vellimedu
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Abstract
A novel process for the purification of recombinant protein expressed as particle is herewith described. In this purification process, the protein is purified by hydrophobic interaction. The interaction of this protein step resulted in an increase in recovery and purity from 15%-80%. The protein further purified has its application in vaccines and pharmaceuticals.
Claims2
A process of purification of a protein of interest from an insoluble fraction, the process consisting of the steps:
- (a) forming an insoluble matrix by the addition of divalent ionic salt having a concentration ranging from 0.2% to 10% with counter ions of either phosphate, chloride and/or acetate solution to the insoluble fraction;
- (b) subjecting the insoluble matrix obtained in step (a) to centrifugation to form a pellet;
- (c) repeatedly subjecting the pellet from step (b) to a desorption process to release said protein(s) from said insoluble pellet by using either TRIS buffer of pH 8.0 to 8.5 or TRIS with EDTA buffer at pH 7.0 to 8.0 to obtain a desorbate;
- (d) recovering said protein from the desorbate through ultrafiltration, chromatography on colloidal silica, hydrophobic chromatography, affinity chromatography, ion exchange, diafiltration, sterile filtration or a combination thereof.
1 more element
Dependencies are inferred from the claim text.