title: Triple-site antigen capture ELISA for human myoglobin can be more effective than double-site assay.
abstract: Using a panel of monoclonal antibodies against human myoglobin (Mb), we have shown that the sensitivity of antigen-capture ELISA can be significantly increased by simultaneous immobilization of two cooperating capture monoclonal antibodies on a solid phase. This method ("triple-site ELISA") uses three monoclonal antibodies to different epitopes of the same antigen (two capture/one tracer) unlike the traditional double-site assay using one capture and one tracer monoclonal antibody. We developed double- and triple-site ELISA for Mb by varying the capture and tracer monoclonal antibodies. Triple-site assays showed 4-6-fold increase in sensitivity compared to the double-site assays. A model for this effect is suggested; according to the model, in triple-site ELISA, high-affinity cyclic configurations can be formed by an antigen, two capture monoclonal antibodies, and the surface of the solid phase.
authors.affiliation: Mental Health Center, Russian Academy of Medical Sciences, Moscow, 113152, Russia.
authors.forename: V A
authors.initials: VA
authors.lastname: Nikulina
authors.forename: E A
authors.initials: EA
authors.lastname: Kizim
authors.forename: Y S
authors.initials: YS
authors.lastname: Massino
authors.forename: O L
authors.initials: OL
authors.lastname: Segal
authors.forename: M B
authors.initials: MB
authors.lastname: Smirnova
authors.forename: V V
authors.initials: VV
authors.lastname: Avilov
authors.forename: D B
authors.initials: DB
authors.lastname: Saprygin
authors.forename: S P
authors.initials: SP
authors.lastname: Smotrov
authors.forename: G I
authors.initials: GI
authors.lastname: Kolyaskina
authors.forename: A D
authors.initials: AD
authors.lastname: Dmitriev
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D006825:Hybridomas; D051379:Mice; D009211:Myoglobin / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D009211:Myoglobin