Synergistic effects in antigen capture ELISA using three monoclonal antibodies directed at different epitopes of the same antigen.
Using a panel of monoclonal antibodies (mAb) against human myoglobin (Mb), we have shown that the sensitivity of antigen-capture enzyme-linked immunosorbent assay (ELISA) may be significantly increased by the simultaneous immobilization on a solid phase of two co-operating capture mAbs. This method ("a three-site ELISA") uses three mAbs at different epitopes of the same antigen (two capture/one tracer), unlike the traditional two-site assay, using one capture and one tracer mAbs. We established two-site and three-site ELISA assays for Mb, by varying capture and tracer mAbs. Three-site assays showed 4-6 fold increase in sensitivity, if compared with two-site assays. The model for the effect has been suggested, according to which in three-site ELISA the high-affinity cyclic configurations may be formed by an antigen, two-capture mAbs and the surface of solid phase.
Nikulina
V A
VA
The Mental Health Center of the Russian Academy of Medical Sciences, Moscow, Russia.
Kizim
E A
EA
Massino
Y S
YS
Segal
O L
OL
Smirnova
M B
MB
Avilov
V V
VV
Saprigin
D B
DB
Smotrov
S P
SP
Tichtchenko
V A
VA
Kolyaskina
G I
GI
Dmitriev
A D
AD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry*; D002138:Calibration; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009206:Myocardium / Q000737:chemistry; D009211:Myoglobin / Q000276:immunology; D011506:Proteins / Q000737:chemistry; D011817:Rabbits; D012015:Reference Standards
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D009211:Myoglobin; D011506:Proteins; D006735:Horseradish Peroxidase