Discrimination by rabbit anti-idiotypic antibodies of two murine IgM monoclonal antibodies directed against lipid A.
Two murine IgM monoclonal antibodies (MAs) directed against the lipid A portion of bacterial lipopolysaccharide (LPS) were compared in their binding to Re LPS and lipid A and their idiotypic make-up with rabbit anti-idiotypic sera. Horseradish peroxidase (HRPO)-labelled MAs 8-2 and 26-20 bound equally well to Re LPS. The binding of HRPO-labelled MA 8-2 to synthetic lipid A was low compared to the relatively strong binding of labelled 26-20. The MAs proved to be competitive in a competition binding assay (CBA) with Re LPS as coating antigen. Rabbit immune sera were raised against individual MAs. Anti-idiotypic antibodies (anti-id Abs) were detected with two sensitive enzyme immunoassays (EIA): a solid-phase EIA and an inhibition EIA. The rabbit antisera proved to be idiotype specific, indicating that both MAs recognize separate epitopes. We expect that anti-id Abs will prove to be of value for the differentiation of panels of LPS specific MAs.
Eijkman-Winkler Laboratory for Microbiology, University Hospital Utrecht, The Netherlands.
J J
JJ
Cornelissen
T A
TA
Oosterlaken
J
J
Schellekens
R
R
Torensma
M
M
Rozenberg-Arska
C A
CA
Kraaijeveld
J
J
Verhoef
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000939:Epitopes; D004926:Escherichia coli / Q000276:immunology; D007124:Immunoenzyme Techniques; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology*; D008050:Lipid A / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D011817:Rabbits; D012475:Salmonella / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007130:Immunoglobulin Idiotypes; D007075:Immunoglobulin M; D008050:Lipid A; D008070:Lipopolysaccharides