Abstract
No other development has had such a major impact on the measurement of hormones as immunoassay. Reagents and assay kits can now be bought commercially but not for the more esoteric or new hormones.
This chapter explains the basics of the immunoassay reaction and gives simple methods for immunoassays and immunometric assays and for the production of reagents for both antigenic and hapten hormones. Alternative methods are given for the preparation of labeled hormones as well as several possible separation procedures. The methods described here have been previously used in a wide range of assays and have stood the test of time. They will allow the production of usable immunoassays in a relatively short period of time.
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References
Yalow RS, Berson SA (1959) Assay of plasma insulin in human subjects by immunological methods. Nature 184:1648–1649
Ekins RP (1960) The estimation of thyroxine in human plasma by an electrophoretic technique. Clin Chim Acta 5:453–459
Rezaieyazdi Z, Hesamifard A (2006) Correlation between serum prolactin and lupus activity. Rheumatol Int 26:1036–1039
Shimizu A, Ohkubo M, Hamaguchi M (2012) Development of non-competitive enzyme-linked immunosorbent assays for mummichog Fundulus heteroclitus gonadotrophins—examining seasonal variations in plasma FSH and LH levels in both sexes. Gen Comp Endocrinol 178:463–469
Sekiguchi S, Kohno H, Yasukawa K et al (2011) Chemiluminescent enzyme immunoassay for measuring leptin. Biosci Biotechnol Biochem 75:752–756
Sun M, Du L, Gao S et al (2010) Determination of 17-beta-estradiol by fluorescence immunoassay with streptavidin-conjugated quantum dots as label. Steroids 75:400–403
Johnsen AH, Assaad FN, Rehfeld JF (2011) Competitive solid-phase immunoassay of gastrin in serum using time-resolved fluorometry. Scand J Clin Lab Invest 71:216–220
Hoferi M, Krist S, Buchbauer G (2005) Adaptation of DELFIA(TM) cortisol kit for determination of salivary cortisol concentration. Arch Pharm 338:493–497
Harlow E, Lane D (eds) (1988) Antibodies: a laboratory manual. Cold Harbor Spring Laboratory, New York, NY
Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of pre-defined specificity. Nature 256:495–497
Zola H (ed) (2000) Monoclonal antibodies. BIOS Scientific Publishers Ltd, Oxford
Allman BL, Short F, James VHT (1981) Fluoroimmunoassay of progesterone in human serum or plasma. Clin Chem 27:1176–1179
Kirkam KE, Hunter WM (eds) (1971) Radioimmunoassay methods. Churchill Livingstone, Edinburgh
Hunter WM, Corrie JET (eds) (1983) Immunoassays for clinical chemistry. Churchill Livingstone, Edinburgh
Avrameas S (1969) Coupling of enzymes to proteins with glutaraldehyde. Immunochemistry 69:43–52
Avrameas S, Ternynck T (1974) Peroxidase labelled antibody and Fab conjugates with enhanced cellular penetration. Immunochemistry 8:1175–1179
Sheng J-W, He M, Shi H-C et al (2006) A comprehensive immunoassay for the detection of microcystins in waters based on polyclonal antibodies. Anal Chim Acta 572:309–315
Ray S, Chowdry P, Das N et al (2010) Development of an efficient and simple method for conjugation of laccase to immunoglobulin and its characterization by enzyme immunoassay. J Immunoassay Immunochem 31:217–232
Wheeler MJ, Barnard G (2010) Immunoassay of steroids. In: Makin HJL, Gower DB (eds) Steroid analysis, 2nd edn. Springer, London, pp 283–327
Wild D (ed) (2013) The immunoassay handbook. Nature Publishing Group, London
Price CP, Newman DJ (eds) (1997) Principles and practice of immunoassay. Stockton, New York
Nars PW, Hunter WM (1973) A method for labelling oestradiol-17β with radioiodine for radioimmunoassay. J Endocrinol 57:157–158
Newman DJ, Price CP (1997) Separation techniques. In: Price CP, Newman DJ (eds) Principles and Practice of immunoassay, 2nd edn. Stockton Press: New York
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Wheeler, M.J. (2013). Immunoassay Techniques. In: Wheeler, M. (eds) Hormone Assays in Biological Fluids. Methods in Molecular Biology, vol 1065. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-616-0_2
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DOI: https://doi.org/10.1007/978-1-62703-616-0_2
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