REMOVAL OF LPS FROM A BRUCELLA CYTOPLASMIC FRACTION BY AFFINITY-CHROMATOGRAPHY WITH AN ANTI-LPS MONOCLONAL-ANTIBODY AS IMMUNOSORBENT

被引:14
作者
GOLDBAUM, FA [1 ]
RUBBI, CP [1 ]
FOSSATI, CA [1 ]
机构
[1] UBA,CONICET,FAC FARM & BIOQUIM,INST ESTUDIOS INMUNIDAD HUMORAL,RA-1113 BUENOS AIRES,DF,ARGENTINA
关键词
D O I
10.1099/00222615-40-3-174
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Affinity chromatography on polymyxin B-Sepharose 4B is one of the most commonly used methods for the removal of contaminating lipopolysaccharides (LPS). However, the LPS of Brucella spp. do not bind to polymyxin B. An affinity chromatography method with an anti-O antigen of Brucella LPS monoclonal antibody as immunosorbent was developed. The method produced a 1000-fold reduction in the LPS content of the cytoplasmic fraction of B. abortus. The eluted proteins retained their antigenicity. The method, which uses mild physiological conditions, is simple, effective and reproducible.
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收藏
页码:174 / 178
页数:5
相关论文
共 23 条
[1]   IMMUNE-RESPONSE OF CATTLE TO BRUCELLA-ABORTUS OUTER-MEMBRANE PROTEINS MEASURED BY LYMPHOCYTE BLASTOGENESIS [J].
BALDWIN, CL ;
VERSTREATE, DR ;
WINTER, AJ .
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 1985, 9 (04) :383-396
[2]   CLONING, EXPRESSION, AND OCCURRENCE OF THE BRUCELLA CU-ZN SUPEROXIDE-DISMUTASE [J].
BRICKER, BJ ;
TABATABAI, LB ;
JUDGE, BA ;
DEYOE, BL ;
MAYFIELD, JE .
INFECTION AND IMMUNITY, 1990, 58 (09) :2935-2939
[3]  
Corbel M. J., 1985, Veterinary Bulletin, V55, P927
[4]  
Galfre G, 1981, Methods Enzymol, V73, P3
[5]   DIFFERENTIATION BETWEEN ACTIVE AND INACTIVE HUMAN BRUCELLOSIS BY MEASURING ANTIPROTEIN HUMORAL IMMUNE-RESPONSES [J].
GOLDBAUM, FA ;
RUBBI, CP ;
WALLACH, JC ;
MIGUEL, SE ;
BALDI, PC ;
FOSSATI, CA .
JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (03) :604-607
[6]   REMOVAL OF GRAM-NEGATIVE ENDOTOXIN FROM SOLUTIONS BY AFFINITY-CHROMATOGRAPHY [J].
ISSEKUTZ, AC .
JOURNAL OF IMMUNOLOGICAL METHODS, 1983, 61 (03) :275-281
[7]   A NEW METHOD FOR REDUCTION OF ENDOTOXIN CONTAMINATION FROM PROTEIN SOLUTIONS [J].
KARPLUS, TE ;
ULEVITCH, RJ ;
WILSON, CB .
JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 105 (02) :211-220
[8]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[9]   CHARACTERIZATION OF THE HEAT-SHOCK RESPONSE IN BRUCELLA-ABORTUS AND ISOLATION OF THE GENES ENCODING THE GROE HEAT-SHOCK PROTEINS [J].
LIN, JS ;
ADAMS, LG ;
FICHT, TA .
INFECTION AND IMMUNITY, 1992, 60 (06) :2425-2431
[10]  
MAYER H, 1984, HDB ENDOTOXIN, V1, P221