Comparison of agarose and dextran-grafted agarose strong ion exchangers for the separation of protein aggregates.
The control of aggregate levels in recombinant protein based drugs is a primary concern during process development and manufacture. In recent years, a novel class of dextran-grafted ion exchange matrices has gained popularity for process scale protein purification due to increased mass transfer rates and higher dynamic binding capacity compared to conventional matrices. Using bovine serum albumin and a monoclonal antibody as model proteins, we studied Sepharose FF and Sepharose XL ion exchangers for the separation of protein aggregates. Experimental results comparing linear gradient elution, stepwise elution, and flow-through chromatography for aggregate separation are described. Differences in performance for the various ion exchangers are discussed and modeled. Strategies for the optimization of protein aggregate separation are provided.
Suda
Eric J
EJ
Pfizer Global Biologics, 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA.
Thomas
Kristin E
KE
Pabst
Timothy M
TM
Mensah
Paul
P
Ramasubramanyan
Natraj
N
Gustafson
Mark E
ME
Hunter
Alan K
AK
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D003911:Dextrans / Q000737:chemistry*; D007475:Ion Exchange Resins / Q000737:chemistry*; D011485:Protein Binding; D012685:Sepharose / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification*
D003911:Dextrans; D007475:Ion Exchange Resins; D012710:Serum Albumin, Bovine; D012685:Sepharose