Performance comparison of Protein A affinity-chromatography sorbents for purifying recombinant monoclonal antibodies

被引:2
作者
Fahrner, RL
Whitney, DH
Vanderlaan, M
Blank, GS
机构
[1] Genentech Inc, Dept Recovery Sci, S San Francisco, CA 94080 USA
[2] Persept Biosyst, POROS Res & Dev, Framingham, MA 01701 USA
[3] Genentech Inc, Dept Analyt Chem, S San Francisco, CA 94080 USA
关键词
D O I
暂无
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页码:121 / 128
页数:8
相关论文
共 33 条
[1]   FLOW-THROUGH PARTICLES FOR THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATION OF BIOMOLECULES - PERFUSION CHROMATOGRAPHY [J].
AFEYAN, NB ;
GORDON, NF ;
MAZSAROFF, I ;
VARADY, L ;
FULTON, SP ;
YANG, YB ;
REGNIER, FE .
JOURNAL OF CHROMATOGRAPHY, 1990, 519 (01) :1-29
[2]   Targeted anti-cancer therapy using rituximab, a chimaeric anti-CD20 antibody (IDEC-C2B8) in the treatment of non-Hodgkin's B-cell lymphoma [J].
Anderson, DR ;
GrilloLopez, A ;
Varns, C ;
Chambers, KS ;
Hanna, N .
BIOCHEMICAL SOCIETY TRANSACTIONS, 1997, 25 (02) :705-708
[3]   Phase II study of weekly intravenous recombinant humanized Anti-p185(HER2) monoclonal antibody in patients with HER2/neu-overexpressing metastatic breast [J].
Baselga, J ;
Tripathy, D ;
Mendelsohn, J ;
Baughman, S ;
Benz, CC ;
Dantis, L ;
Sklarin, NT ;
Seidman, AD ;
Hudis, CA ;
Moore, J ;
Rosen, PP ;
Twaddell, T ;
Henderson, IC ;
Norton, L .
JOURNAL OF CLINICAL ONCOLOGY, 1996, 14 (03) :737-744
[4]  
Bodey B, 1996, ANTICANCER RES, V16, P517
[5]   HUMANIZATION OF AN ANTI-P185HER2 ANTIBODY FOR HUMAN CANCER-THERAPY [J].
CARTER, P ;
PRESTA, L ;
GORMAN, CM ;
RIDGWAY, JBB ;
HENNER, D ;
WONG, WLT ;
ROWLAND, AM ;
KOTTS, C ;
CARVER, ME ;
SHEPARD, HM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (10) :4285-4289
[6]   AFFINITY PURIFICATION OF PROTEINS USING EXPANDED BEDS [J].
CHASE, HA ;
DRAEGER, NM .
JOURNAL OF CHROMATOGRAPHY, 1992, 597 (1-2) :129-145
[7]  
CHEN AB, 1996, J BIOTECHNOL HEALTHC, V3, P70
[8]   ISOLATION OF PURE IGG1, IGG2A AND IGG2B IMMUNOGLOBULINS FROM MOUSE SERUM USING PROTEIN A-SEPHAROSE [J].
EY, PL ;
PROWSE, SJ ;
JENKIN, CR .
IMMUNOCHEMISTRY, 1978, 15 (07) :429-436
[9]  
FAHMER RL, 1999, BIOTECHNOL APPL BIOC, V29, P109
[10]   OPTIMIZATION OF THE EXPERIMENTAL CONDITIONS AND THE COLUMN DESIGN PARAMETERS IN OVERLOADED ELUTION CHROMATOGRAPHY [J].
FELINGER, A ;
GUIOCHON, G .
JOURNAL OF CHROMATOGRAPHY, 1992, 591 (1-2) :31-45