NEUTRALIZING MONOCLONAL-ANTIBODIES TO PASTEURELLA-HAEMOLYTICA LEUKOTOXIN AFFINITY-PURIFY THE TOXIN FROM CRUDE CULTURE SUPERNATANTS

被引:76
作者
GENTRY, MJ [1 ]
SRIKUMARAN, S [1 ]
机构
[1] UNIV NEBRASKA,DEPT VET SCI,LINCOLN,NE 68583
关键词
PASTEURELLA-HAEMOLYTICA; LEUKOTOXIN NEUTRALIZATION; AFFINITY-PURIFICATION; MONOCLONAL ANTIBODY;
D O I
10.1016/0882-4010(91)90086-P
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The leukotoxin of Pasteurella haemolytica is a major virulence factor of the organism. It is an unstable protein which has proven very difficult to purify using traditional techniques. Hybridomas secreting monoclonal antibodies (mAbs) to P. haemolytica leukotoxin were derived from spleen cells of a mouse immunized with crude culture supernatant. Five hybridomas secreting mAbs specific for the leukotoxin were stabilized. Each of the mAbs reacted with a protein of approximately 100 kDa in toxic culture supernatants, and two of them completely neutralized the toxin in vitro. Affinity chromatography of crude culture supernatant on a column prepared with one of the neutralizing mAbs resulted in the isolation of biologically active toxin. © 1991.
引用
收藏
页码:411 / 417
页数:7
相关论文
共 24 条
[1]  
BALUYUT CS, 1981, AM J VET RES, V42, P1920
[2]  
BERGGREN KA, 1981, AM J VET RES, V42, P1383
[3]   CHEMICAL AND IMMUNOLOGICAL ANALYSIS OF THE COMPLEX STRUCTURE OF ESCHERICHIA-COLI ALPHA-HEMOLYSIN [J].
BOHACH, GA ;
SNYDER, IS .
JOURNAL OF BACTERIOLOGY, 1985, 164 (03) :1071-1080
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P47
[5]  
CHANG YF, 1986, AM J VET RES, V47, P67
[6]   IDENTIFICATION AND CHARACTERIZATION OF THE PASTEURELLA-HAEMOLYTICA LEUKOTOXIN [J].
CHANG, YF ;
RY, Y ;
POST, D ;
STRUCK, DK .
INFECTION AND IMMUNITY, 1987, 55 (10) :2348-2354
[7]  
CHANG YF, 1986, AM J VET RES, V47, P716
[8]  
CORSTVET RE, 1973, J AM VET MED ASSOC, V163, P870
[9]   IMMUNOSEROLOGICAL COMPARISON OF 104-KILODALTON PROTEINS ASSOCIATED WITH HEMOLYSIS AND CYTOLYSIS IN ACTINOBACILLUS-PLEUROPNEUMONIAE, ACTINOBACILLUS-SUIS, PASTEURELLA-HAEMOLYTICA, AND ESCHERICHIA-COLI [J].
DEVENISH, J ;
ROSENDAL, S ;
JOHNSON, R ;
HUBLER, S .
INFECTION AND IMMUNITY, 1989, 57 (10) :3210-3213
[10]   COLORIMETRIC METHOD FOR DETERMINATION OF SUGARS AND RELATED SUBSTANCES [J].
DUBOIS, M ;
GILLES, KA ;
HAMILTON, JK ;
REBERS, PA ;
SMITH, F .
ANALYTICAL CHEMISTRY, 1956, 28 (03) :350-356