Abstract
A sensitive immunoenzymometric assay (IEMA) of serum thyrotropin (hTSH) was developed using anti-hTSH rabbit polyclonal antibody and anti-hTSH in-house monoclonal antibody with a sensitivity of 0.12 mIU/L. Serum samples were incubated in ELISA wells precoated with polyclonal antibody. The hTSH bound to the wells was incubated with monoclonal antibody (detector antibody) and further with goat anti-mouse antibody–horse radish peroxidase (GAM-HRP), which obviates the need to label the detector antibody. The assay was validated by recovery, linearity, and cross-reactivity experiments with a working assay range of 0.15 to 100 mIU/L and <10% coefficient of variation (CV) for both intra- and interassay. Good correlations were obtained when compared with Immunotech hTSH IRMA (r = 0.971, n = 35). This in-house ELISA can be used as an initial screening test for thyroid dysfunction.
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