Application of high-performance tangential flow filtration (HPTFF) to the purification of a human pharmaceutical antibody fragment expressed in Escherichia coli.
High-performance tangential flow filtration (HPTFF) is shown to successfully enable concentration, purification and formulation in a single unit operation. This is illustrated with feedstreams comprising recombinant proteins expressed in Escherichia coli (E. coli). Using positively charged cellulosic membranes of 100 kDa molecular weight cut-off and operating under a selected range of buffer pH and ionic strength at a filtrate flux of 100 L m(-2) h(-1), a 10-fold removal of E. coli host cell proteins (HCP) was obtained with an overall process yield of 98%. The HPTFF performance was shown to be robust and reproducible. In addition, the novel charged membrane was regenerated and re-used seven times without loss of selectivity or throughput. When compared with a conventional purification scheme, the proposed process results in the elimination of one chromatographic step, a 12% yield improvement and a significant reduction in purification cost of goods.
Lebreton
Bénédicte
B
Late Stage Purification, Process Development, Genentech Inc., 1 DNA way, South San Francisco, California 94080, USA. lebreton.benedicte.@gene.com
Brown
Arick
A
van Reis
Robert
R
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D004364:Pharmaceutical Preparations / Q000302:isolation & purification*; D014462:Ultrafiltration / Q000295:instrumentation* / Q000379:methods
D000911:Antibodies, Monoclonal; D004364:Pharmaceutical Preparations