Expanded bed adsorption in the purification of monoclonal antibodies: a comparison of process alternatives

被引:27
作者
Blank, GS [1 ]
Zapata, G [1 ]
Fahrner, R [1 ]
Milton, M [1 ]
Yedinak, C [1 ]
Knudsen, H [1 ]
Schmelzer, C [1 ]
机构
[1] Genentech Inc, Dept Recovery Sci, San Francisco, CA 94080 USA
关键词
expanded bed; Protein A; cation exchange; monoclonal antibodies;
D O I
10.1023/A:1012278022349
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Expanded bed adsorption (EBA) was examined as the initial capture/purification step in the purification of monoclonal antibodies from Chinese hamster ovary (CHO) cultures. Two process alternatives each using EBA were compared to a conventional Protein A process without EBA. One alternative used Protein A affinity EBA followed by packed-bed cation and anion-exchange steps. The other alternative used cation-exchange EBA as the capture step followed by packed-bed Protein A and anion-exchange steps. The process using Protein A EBA produced comparable purity (host cell protein, DNA, Protein A, antibody aggregate) to the conventional process. However, the Protein A EBA column showed a significant decrease in dynamic capacity with a limited number of cycles. The process using cation EBA achieved comparable levels of host cell proteins (HCP) and DNA but not antibody aggregate or leached Protein A compared to the conventional process.
引用
收藏
页码:65 / 71
页数:7
相关论文
共 7 条
[1]  
ERICKSON JC, 1994, ANIMAL CELL TECHNOLO, P557
[2]   Expanded bed protein A affinity chromatography of a recombinant humanized monoclonal antibody: process development, operation, and comparison with a packed bed method [J].
Fahrner, RL ;
Blank, GS ;
Zapata, GA .
JOURNAL OF BIOTECHNOLOGY, 1999, 75 (2-3) :273-280
[3]   PILOT-SCALE RECOVERY OF RECOMBINANT ANNEXIN-V FROM UNCLARIFIED ESCHERICHIA-COLI HOMOGENATE USING EXPANDED BED ADSORPTION [J].
FREJ, AKB ;
HJORTH, R ;
HAMMARSTROM, A .
BIOTECHNOLOGY AND BIOENGINEERING, 1994, 44 (08) :922-929
[4]   SINGLE-STEP RECOVERY OF A SECRETED RECOMBINANT PROTEIN BY EXPANDED BED ADSORPTION [J].
HANSSON, M ;
STAHL, S ;
HJORTH, R ;
UHLEN, M ;
MOKS, T .
BIO-TECHNOLOGY, 1994, 12 (03) :285-288
[5]   Isolation of monoclonal antibodies from cell containing hybridoma broth using a protein A coated adsorbent in expanded beds [J].
Thommes, J ;
Bader, A ;
Halfar, M ;
Karau, A ;
Kula, MR .
JOURNAL OF CHROMATOGRAPHY A, 1996, 752 (1-2) :111-122
[6]   PURIFICATION OF MONOCLONAL-ANTIBODIES FROM WHOLE HYBRIDOMA FERMENTATION BROTH BY FLUIDIZED-BED ADSORPTION [J].
THOMMES, J ;
HALFAR, M ;
LENZ, S ;
KULA, MR .
BIOTECHNOLOGY AND BIOENGINEERING, 1995, 45 (03) :205-211
[7]  
ZAPATA G, 1999, BIOTHERAPEUTICS 99 E