Interference of complement with the binding of carcinoembryonic antigen to solid-phase monoclonal antibodies.
Six mouse monoclonal antibodies against carcinoembryonic antigen were evaluated for use in a solid-phase immunometric assay. Three IgG2 antibodies, when attached to polymer particles or microtiter wells, were found to be severely inhibited by fresh serum. The remaining three antibodies, which were of the IgG1 subclass, were inhibited only slightly or not at all when used in the same way. With the aid of labelled antibodies against C1q and C3, it was shown that antibody inhibition was accompanied by the binding of large amounts of these complement factors. Experiments involving heat inactivation or dilutions of serum, or the addition of EDTA, consistently revealed the same correlation between binding of complement factors and inhibition of CEA binding to antibody. It is suggested that the significant inhibition of the solid-phase IgG2 antibodies was caused by classical pathway activation of complement by the solid-phase antibody even in the absence of antigen. The slight inhibition of solid-phase IgG1 antibodies observed at high serum concentrations was probably due to complement binding by the alternative pathway. Preliminary evidence suggests that complement interference is not restricted to CEA assays, but is a potential problem in all types of serum immunoassays using solid-phase antibodies.
Börmer
O P
OP
Central Laboratory, Norwegian Radium Hospital, Oslo.
D000818:Animals; D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen / Q000032:analysis* / Q000276:immunology; D003167:Complement Activation; D003165:Complement System Proteins / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D007118:Immunoassay; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D007074:Immunoglobulin G; D003165:Complement System Proteins; D004492:Edetic Acid