Rapid and simple measurement of human C5a-des-Arg level in plasma or serum using monoclonal antibodies.
A new sandwich immunoassay method for measuring human C5a-des-Arg was developed using monoclonal antibody specifically reactive with C5a-des-Arg. Monoclonal antibodies were obtained from a panel of hybridomas produced by fusion of mouse myeloma cells, P3 X 63-AG8,653, with spleen cells from a CBF1(C57BL/6 X BALB/c) mouse immunized with purified C5a. The reactivities of these monoclonal antibodies against C5a, C5a-des-Arg and C5 were tested by solid-phase radioimmunoassay. One of the antibodies reacted with C5a-des-Arg, but not with C5a and C5. By use of this antibody for capturing antibody in sandwich immunoassay, a rapid and simple method was developed for measuring C5a-des-Arg without previous removal of C5. The sensitivity of this assay system was approximately 1 ng/ml for C5a-des-Arg.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003182:Complement C5 / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D015936:Complement C5a; D015937:Complement C5a, des-Arginine; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D003182:Complement C5; D015937:Complement C5a, des-Arginine; D015936:Complement C5a