[Mathematical analysis of equilibrium on 2-site immunoradiometric assay using anti-human TSH monoclonal antibody].
A mathematical analysis of equilibrium was adapted on a practical 2-site immunoradiometric assay (IRMA) using 10 murine monoclonal antibodies (Mabs) to human TSH. The affinity constant of 10 Mabs was from 2.5 x 10(8) to 3.3 x 10(10) M-1. The dose-response curves using appropriate combinations of Mab-coated bead and 125I-labeled Mab were compared with the theoretical curves calculated from mathematical analysis of equilibrium. Theoretical curves were directly affected by the affinity constants in which antibodies with higher affinity constants showed higher binding activity of tracer. However, the curves which antibodies have more than 1 x 10(9) M-1 and 1 x 10(10) M-1 for capture antibody and tracer antibody did not show remarkable change, but were similar with the curve obtained from unlimited affinity constant. From these curve, the maximum and minimum detectable dose were approx. 2 x 10(-10) M and 4 x 10(-14) M, respectively. Seven out of 10 experimental curves for TSH showed good consistency with the corresponding theoretical curves which indicated that a mathematical analysis of equilibrium was useful to presume experimental results using monoclonal antibody.
Obata
K
K
Kubo
Y
Y
Okane
Y
Y
Kato
A
A
Ikeda
I
I
Kurata
K
K
D000911:Antibodies, Monoclonal; D006801:Humans; D008433:Mathematics; D011863:Radioimmunoassay / Q000379:methods*; D011933:Reagent Kits, Diagnostic / Q000592:standards; D013972:Thyrotropin / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D011933:Reagent Kits, Diagnostic; D013972:Thyrotropin