Performance comparison of protein A affinity-chromatography sorbents for purifying recombinant monoclonal antibodies.
We describe the performance characteristics of five Protein A affinity-chromatography sorbents (Sepharose Fast Flow, Poros 50, Poros LP, Prosep and Streamline) for purifying a recombinant humanized monoclonal antibody from clarified Chinese hamster ovary cell culture fluid. We measured the dynamic capacity at varying flow rates, maximum capacity, pressure drop and production rate. For purified antibody, we measured yield and purity (by SDS/PAGE, the amount of DNA, the amount of host-cell proteins and the amount of Protein A). We found that, whereas all sorbents provided significant and essentially equivalent antibody purification, there were differences in capacity and pressure drop, which affected the production rate and had implications for process applications.
Fahrner
R L
RL
Department of Recovery Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Whitney
D H
DH
Vanderlaan
M
M
Blank
G S
GS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000378:metabolism; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002847:Chromatography, Agarose / Q000295:instrumentation / Q000379:methods; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A