Enzyme immunoassay of thyrotropin releasing hormone (TRH).
A sensitive and specific double-antibody enzyme immunoassay (EIA) for a thyrotropin releasing hormone (TRH)-like immunoreactive substance has been developed. In order to synthesize TRH-labeled beta-D-galactosidase (beta-gal), a newly devised TRH derivative, pGlu-His-Pro-NH-(CH2)6-NH2 (TRH-Hex), was employed. TRH-Hex was linked to beta-gal by the N-(epsilon-maleimidocaproyloxy) succinimide coupling procedure. For competitive reactions, the TRH antibody was incubated with standard TRH and TRH-Hex-beta-gal (delayed addition). Free and antibody-bound enzyme hapten were separated by using an anti-rabbit immunoglobulin G coated immunoplate. Activity of the enzyme on the plate was fluorometrically determined. The present immunoassay allows detection of 0.8 to 100 pmol/well of TRH.
Takeyama
M
M
Department of Clinical Pharmacy, Oita Medical University, Japan.
Otaka
A
A
Fujii
N
N
D000595:Amino Acid Sequence; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D013973:Thyrotropin-Releasing Hormone / Q000032:analysis* / Q000276:immunology
D013973:Thyrotropin-Releasing Hormone