A sensitive enzyme immunoassay for the quantitation of human C5a/C5a(desArg) anaphylatoxin using a monoclonal antibody with specificity for a neoepitope.
The direct quantitation of C5a/C5a(desArg) in human plasma was achieved by a specific and highly sensitive ELISA which is based on the neoepitope-specific monoclonal antibody C17/5. The error-prone removal of C5 from plasma prior to the assay is therefore not required. With a detection limit of 20 pg C5a/ml plasma, the sensitivity of this assay allowed to define normal ranges (1.94 +/- 1.49 ng C5a/nl, mean +/- SD) of C5a/C5a(desArg) in human blood plasma. This assay will also be applicable to the quantitation of C5a in specimens with low protein content where precipitation-based assays fail to accurately determine C5a. In addition, the mAb C17/5 turned out to efficiently block the binding of C5a/C5a (desArg) to its cellular receptor and therefore provides a valuable tool in the delineation of C5a effects in complex biologic systems in the presence of native C5, such as under in vivo conditions.
Oppermann
M
M
Department of Immunology, University of Göttingen, FRG.
Schulze
M
M
Götze
O
O
D000255:Adenosine Triphosphate / Q000097:blood; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity / Q000276:immunology; D001782:Blood Donors; D001792:Blood Platelets / Q000378:metabolism; D001793:Blood Preservation; D002138:Calibration; D015936:Complement C5a / Q000032:analysis* / Q000276:immunology; D015937:Complement C5a, des-Arginine / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D012680:Sensitivity and Specificity; D013696:Temperature
D000911:Antibodies, Monoclonal; D015937:Complement C5a, des-Arginine; D000939:Epitopes; D015936:Complement C5a; D000255:Adenosine Triphosphate