The effect of antigen cross-linking on the sensitivity of the enzyme-linked immunosorbent assay.
To investigate the effects of antigen cross-linking on ELISA results, we assayed antibodies to several purified antigens, using microtitration trays coated with antigens in their native and cross-linked forms. Cross-linking was achieved by a carbodiimide promoted condensation reaction. For several antigens an increase in sensitivity of the ELISA was obtained after cross-linking. The results of a double antibody sandwich assay using the same cross-linked antigens, showed that the affinity for antibodies remained unchanged after limited cross-linking. Therefore, the higher sensitivity of the ELISA is most likely due to a higher affinity of the cross-linked antigens for the plastic carrier. Cross-linking of proteins in a complex antigen preparation obtained from Schistosoma mansoni led to a large increase in sensitivity.
D000818:Animals; D000941:Antigens / Q000276:immunology; D002234:Carbodiimides / Q000494:pharmacology*; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000378:metabolism; D046911:Macromolecular Substances; D010047:Ovalbumin / Q000276:immunology / Q000378:metabolism; D012550:Schistosoma mansoni / Q000276:immunology; D012552:Schistosomiasis / Q000276:immunology
D000941:Antigens; D002234:Carbodiimides; D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D010047:Ovalbumin