Routine manufacture of recombinant proteins using expanded bed adsorption chromatography

• Published in: Bioseparation Vol. 10; no. 1-3; p. 51
• Main Authors: Shepard, S R, Boyd, G A, Schrimsher, J L
• Format: Journal Article
• Language: English
• Published: Netherlands 2001
• Subjects: Adsorption; Chromatography, Liquid - methods; Electrophoresis, Polyacrylamide Gel; Humans; Pichia - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification
• ISSN: 0923-179X
• DOI: 10.1023/a:1012096020895

Two different recombinant human proteins were purified directly from Pichia pastoris whole cell fermentation broth, containing 30-44% biomass (wet weight percent), by strong cation exchange expanded bed adsorption chromatography. Expanded bed adsorption chromatography provided clarification, product purification and product concentration in a single unit operation at large scale (2000-1 nominal fermentation volume). The efficiency of expanded bed adsorption chromatography resulted in a short process time, high process yield, and limited proteolytic degradation of the target proteins. The separations were operated using a 60-cm (d) column run at 14 l/min. For one protein, expanded bed adsorption chromatography resulted in an average product recovery of 113% (relative to fermentation supernatant) and a purity of 89% (n=10). For the other protein, the average product recovery was 99% (relative to fermentation supernatant) and the purity was 62.1 (n=10). Laboratory experiments showed that biomass reduced product dynamic binding capacity for protein 2.