Enzyme immunometric assay of thyroliberin (TRH).
An enzyme immunometric assay of thyroliberin (TRH) using monoclonal antibodies and a derivatization procedure is described. This assay, named SPIE-IA, involves a four step procedure after chemical derivatization of TRH and biological samples by diazotized APEA. Step 1: derivatized TRH was immunocaptured by a monoclonal anti-TRH antibody coated on a 96-well microtiter plate. Step 2: after washing, derivatized TRH was cross-linked via its amino group to the wells using glutaraldehyde. Step 3: washing and treatment with NaOH. Step 4: measurement of bound TRH using a monoclonal anti-TRH antibody labeled with acetylcholinesterase. The minimal detectable concentration was 0.1 pmol/ml: with a coefficient of variation less than 10% in the 0.156-10 pmol/ml range. This assay is 26-fold more sensitive and more specific than the competitive enzyme immunoassay using the same monoclonal capture antibody, derivatized TRH and TRH-acetylcholinesterase conjugate as tracer. Good correlation was observed between SPIE-IA and a sensitive competitive enzyme immunoassay using polyclonal antibodies.
Etienne
E
E
Commissariat à l'Energie Atomique, Service de Pharmacologie et d'Immunologie, Gif sur Yvette, France.
Créminon
C
C
Grassi
J
J
Grouselle
D
D
Roland
J
J
Pradelles
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001667:Binding, Competitive / Q000276:immunology; D007124:Immunoenzyme Techniques; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013973:Thyrotropin-Releasing Hormone / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D013973:Thyrotropin-Releasing Hormone