Abstract
A procedure for immunoradiometric assay (IRMA) using solid phase Sepharose particles for the measurement of thyroid stimulating hormone (TSH) in human serum or plasma was developed, optimized, and validated. The 125I-anti-TSH monoclonal antibody (MoAb) was prepared using N-bromosuccinimide as oxidizing agent and was purified by HPLC. The coupling of activated sepharose particles with polyclonal antiobody of TSH produced using rabbit as a host animal was undertaken. Optimization and validation of the assay were carried out. The reproducibility as measured by the intra- and interassay variations is acceptable. The recovery and dilution tests confirmed that the calibration was accurate and the matrix was appropriate. The results agree well with those obtained with the currently used commercial kit. The technique developed proved to be sensitive, specific, precise, and accurate for routine laboratory use.
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