REPEATED CLEANING OF PROTEIN-A AFFINITY COLUMN WITH SODIUM-HYDROXIDE

被引:61
作者
HALE, G
DRUMM, A
HARRISON, P
PHILLIPS, J
机构
[1] Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, Tennis Court Road
基金
英国惠康基金;
关键词
PROTEIN A; SODIUM HYDROXIDE; CLEANING; ENDOTOXIN; MONOCLONAL ANTIBODY; CHROMATOGRAPHY; AFFINITY;
D O I
10.1016/0022-1759(94)90223-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Protein A attached to an affinity adsorbent was shown to be remarkably resistant to 0.5 M NaOH. Even repeated treatments gave only a small decrease in functional capacity and no adverse effect on leakage of the protein A into the eluate. This simple cleaning procedure should be useful in applications where antibodies need to be purified free from the risk of contamination with endotoxins or micro-organisms, e.g., for in vivo therapy, either in clinical trials or experimental animals. It can also prevent cross-contamination when the same protein A adsorbent is used for different batches of antibody.
引用
收藏
页码:15 / 21
页数:7
相关论文
共 21 条
[1]   DETECTION AND REDUCTION OF PROTEIN-A CONTAMINATION IN IMMOBILIZED PROTEIN-A PURIFIED MONOCLONAL-ANTIBODY PREPARATIONS [J].
BLOOM, JW ;
WONG, MF ;
MITRA, G .
JOURNAL OF IMMUNOLOGICAL METHODS, 1989, 117 (01) :83-89
[2]  
BROWN TT, 1981, AM J VET RES, V42, P1033
[3]  
CLARK MR, 1986, METHOD ENZYMOL, V121, P548
[4]   ASSAYS USING RED CELL-LABELED ANTIBODIES [J].
COOMBS, RRA ;
SCOTT, ML ;
CRANAGE, MP .
JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 101 (01) :1-14
[5]  
DRULAK MW, 1984, MICROBIOS, V41, P31
[6]   COMPARISON OF IMMUNOGLOBULIN BINDING-CAPACITIES AND LIGAND LEAKAGE USING 8 DIFFERENT PROTEIN-A AFFINITY-CHROMATOGRAPHY MATRICES [J].
FUGLISTALLER, P .
JOURNAL OF IMMUNOLOGICAL METHODS, 1989, 124 (02) :171-177
[7]   COMPOSITE AFFINITY SORBENTS AND THEIR CLEANING IN PLACE [J].
GIROT, P ;
MOROUX, Y ;
DUTEIL, XP ;
NGUYEN, C ;
BOSCHETTI, E .
JOURNAL OF CHROMATOGRAPHY, 1990, 510 :213-223
[8]   A SENSITIVE ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA) FOR THE DETECTION OF STAPHYLOCOCCAL PROTEIN-A (SPA) PRESENT AS A TRACE CONTAMINANT OF MURINE IMMUNOGLOBULINS PURIFIED ON IMMOBILIZED PROTEIN-A [J].
GODFREY, MAJ ;
KWASOWSKI, P ;
CLIFT, R ;
MARKS, V .
JOURNAL OF IMMUNOLOGICAL METHODS, 1992, 149 (01) :21-27
[9]   USE OF STAPHYLOCOCCAL PROTEIN-A AS AN IMMUNOLOGICAL REAGENT [J].
GODING, JW .
JOURNAL OF IMMUNOLOGICAL METHODS, 1978, 20 (APR) :241-253
[10]   ISOLATION OF LOW-FREQUENCY CLASS-SWITCH VARIANTS FROM RAT HYBRID MYELOMAS [J].
HALE, G ;
COBBOLD, SP ;
WALDMANN, H ;
EASTER, G ;
MATEJTSCHUK, P ;
COOMBS, RRA .
JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 103 (01) :59-67