Radiobinding Assay: A Sensitive Approach for Selecting Anti-TSH Antibodies in Thyroid Disorder Detection
Abstract
The study aimed to evaluate the Radiobinding Assay (RBA) as a sensitive method for assessing antibody responses following immunization, and to compare its performance with the conventional enzyme-linked immunosorbent assay (ELISA). BALB/c mice were immunized with purified human thyroid-stimulating hormone (hTSH) using Freund’s adjuvants, and antisera were collected after successive booster injections. Antibody production was initially screened by indirect ELISA and subsequently re-evaluated by RBA employing 125I-labeled hTSH to measure antigen–antibody binding with higher precision. RBA analysis revealed notable inter-individual variability and demonstrated superior sensitivity in differentiating strong from weak immune responders. Among the immunized mice, serum from IM4 exhibited the highest binding activity, reaching 68.2% at 1:300 dilution. These findings highlight the value of RBA as a robust and highly sensitive approach for identifying optimal antibody producers. This approach provides a strong foundation for subsequent monoclonal antibody generation and the development of advanced immunoassays for the clinical detection of thyroid disorders.
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