Rapid, homogeneous phase, liposome-based assays for total complement activity.
A simple, rapid assay for determining total complement activity has been developed. Complement activity is quantitated spectrophotometrically by measuring the amount of liposome-encapsulated enzyme unmasked by the action of anti-Dnp antibody and complement on Dnp-tagged liposomes. The assay is homogeneous in nature and is nonisotopic. The activity of complement in guinea pig serum has been measured and shown to be proportional to complement concentration. The assay was modified to measure the complement-fixing titer of anti-Dnp antibody preparations. We have compared two monoclonal anti-Dnp antibodies (IgG1 and IgM) for their ability to fix complement. The IgM antibody preparation was 450-fold more effective than the IgG1 preparation in mediating complement-dependent damage to Dnp liposomes. In addition, the test was modified to measure complement fixation by soluble antigen-antibody complexes. This complement fixation format is capable of detecting 2 pmol Dnp antigen.
D019297:2,4-Dinitrophenol; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D000941:Antigens / Q000276:immunology; D003168:Complement Fixation Tests; D003165:Complement System Proteins / Q000032:analysis*; D004140:Dinitrophenols / Q000276:immunology; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D008081:Liposomes
D000911:Antibodies, Monoclonal; D000941:Antigens; D004140:Dinitrophenols; D008081:Liposomes; D003165:Complement System Proteins; D019297:2,4-Dinitrophenol