Production and characterization of monoclonal and polyclonal antibodies against thyrotropin-releasing hormone.

ABSTRACT

In this article, two mouse monoclonal antibodies (83-7B5-A1 and 83-6B6-A10) and three rabbit polyclonal antibodies (1118, 8572, and 8577) directed against thyrotropin-releasing hormone (TRH; pGlu-His-Pro-NH2) are described. The anti-TRH antibodies were raised by immunization with a TRH-bovine serum albumin conjugate obtained by coupling of the CO2H group of pGlu-His-Pro-OH to NH2 groups in the protein. The monoclonal antibodies were produced by hybridoma clones obtained by the fusion of SP2/0 myeloma cells with spleen cells of an immunized BALB/c mouse. Both monoclonal antibodies were of the IgG1 (kappa) subclass. Characterization of the anti-TRH antibodies showed that in general they are specific for the pGlu-His moiety. The cross-reactivities for the TRH-like peptides [Glu1]TRH, [Glu2]TRH, and [Phe2]TRH are low, while alterations at the Pro-NH2 moiety of TRH are recognized to varying extents. The specificities of these antibodies are markedly different from those previously obtained using TRH coupled through the histidine residue to protein as the immunogen.