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Development of solid support using protein A for the measurement of free thyroxine in human serum

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Abstract

Protein A is a 56 kDa cell wall component of Staphylococcus aureus. It binds to the Fc part of the antibody with high affinity. Therefore, it can be used for the purification of antibodies in a plethora of applications. Additionally, it was applied for the immobilization of antibodies to solid supports. Herein, the development of free thyroxine radioimmunoassay (FT4 RIA) using protein A for immobilizing anti-T4 antibodies in the polystyrene tubes is described. 250µL of 5 µg/mL solution was sufficient to yield maximum coating efficiency with uniform and stable antibody-coated tubes with very less nonspecific binding. This approach was economical with respect to the consumption of antibodies and the time required for the preparation of antibody-coated tubes. This resulted in a FT4 assay with a sensitivity of 1pg/mL, with intra and inter-assay variation of less than 9.6 and 11.5% respectively, analyzed at three different FT4 concentrations. The developed system correlated well with the established FT4 RIA kit with a correlation coefficient of 0.985.

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Acknowledgements

Authors thank Dr. N. Sivaprasad, Former SGM, RPH, and Dr. Grace Samuel, Former SGM, QC, RPH, BRIT for their keen interest and support in this project.

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  1. Department of Atomic Energy, Board of Radiation and Isotope Technology (BRIT), BARC Vashi Complex, 400703, Navi Mumbai, India

    Rani Gnanasekar, Vishwas Murhekar & V B Kadwad

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  1. Rani Gnanasekar
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  2. Vishwas Murhekar
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  3. V B Kadwad
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Correspondence to Rani Gnanasekar.

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Gnanasekar, R., Murhekar, V. & Kadwad, V.B. Development of solid support using protein A for the measurement of free thyroxine in human serum. J Radioanal Nucl Chem 331, 4135–4139 (2022). https://doi.org/10.1007/s10967-022-08482-z

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