Triple-site antigen capture ELISA for human myoglobin can be more effective than double-site assay.
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.
Nikulina
V A
VA
Mental Health Center, Russian Academy of Medical Sciences, Moscow, 113152, Russia.
Kizim
E A
EA
Massino
Y S
YS
Segal
O L
OL
Smirnova
M B
MB
Avilov
V V
VV
Saprygin
D B
DB
Smotrov
S P
SP
Kolyaskina
G I
GI
Dmitriev
A D
AD
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
D000911:Antibodies, Monoclonal; D000941:Antigens; D009211:Myoglobin