Abstract
We describe a simple method for the immobilisation of anti-thyroxine antibody on to the surface of polystyrene tubes and a simple assay format for the quantitative estimation of total thyroxine in serum. The immobilisation of anti-thyroxine antibody was achieved through passive adsorption of normal rabbit gamma globulin and anti-rabbit antibody raised in goat, as immune bridges. This procedure ensured minimum utilisation of primary and secondary antibody as neat sera without precipitation or affinity purification. The developed assay system using these antibody coated tubes covers a range of 0–240 ng/mL of thyroxine with intra and inter assay variations of less than 10 %.
References
Tijssen P (1985) Practice and theory of enzyme immunoassays. Elsevier, Amsterdam
Butler JE, Ni L, Brown WR et al (1993) The immunochemistry of sandwich ELISAs-VI. Greater than 90% of monoclonal and 75% of polyclonal anti-fluorescyl capture antibodies are denatured by passive adsorption. Mol Immunol 30:1165
Butler JE, Ni L, Nessler R (1992) The physical and functional behaviour of capture antibodies adsorbed on polystyrene. J Immunol Methods 150:77–90
Kakabakos SE, Evangelatos GP, Ithakissios DS (1990) Immunoadsorption of IgG on to second antibody covalently attached to Amino-Dylark beads for radioimmunoassays. Clin Chem 36:497
Peterman JH, Tarcha PJ, Chu VP, Buttler JE (1988) The immunochemistry of sandwich ELISAs IV. A comparison of the antigen capture capacity of antibodies adsorbed or covalently attached to polystyrene. J Immunol Method 11:271
Petrou PS, Kakabakos SE (1998) Antibody coating approach involving gamma globulins from nonimmunized animal and second antibody antiserum. J Immunoass 19:271
Rani Gnanasekar UH, Nagvekar N Sivaprasad (2010) Development and validation of a two-step free thyroxine radioimmunoassay based on antibody coated tubes. J Liq Chromatogr Relat Technol 33(17):1576–1586
Greenwood F, Hunter WM, Glover JS (1963) J Biochem 9:114
Odell WD (1983) Radiolabelling techniques. In: Odell WD, Franchimont P (eds) principles of competitive protein binding assays. Wiley, New York, pp 69–83
Bhupal V, Mani RS (1986) A simple method for the determination of the specific activity of 125Itracer used in radioimmunoassay. J Radioanal Nucl Chem 107(6):377–383
Acknowledgments
The authors thank Dr. A.K. Kohli, Chief Executive, BRIT for his constant support and encouragement. We sincerely thank Late Shri.U.H. Nagvekar who had initiated this work.
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K. Bapat: Office of Principal Scientific Adviser to the Govt. Of India, New Delhi.
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Gnanasekar, R., Paradkar, S., Kadwad, V. et al. Development of a simple method for the immobilization of anti-thyroxine antibody on polystyrene tubes for use in the measurement of total thyroxine in serum. J Radioanal Nucl Chem 303, 2579–2583 (2015). https://doi.org/10.1007/s10967-014-3829-z
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DOI: https://doi.org/10.1007/s10967-014-3829-z