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The Enzyme Linked Immunosorbent Assay (ELISA)

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Techniques in Molecular Biology

Abstract

Enzyme linked immunosorbent assays (ELISA) provide sensitive methods for detecting either antigens or antibodies in biological fluids, and as such have found numerous applications, particularly in the field of clinical analysis but also in applications to research in molecular biology (see below, p. 94). The first ELISA techniques were described by Engvall and Perlmann1 and Van Weeman and Schuurs2 working independently in 1971. The principles of ELISA are similar to those of radioimmunoassay techniques3 except that instead of using a radiolabel-linked antigen or antibody to quantitate antigen-antibody reactions, an enzyme label is used instead. Depending on the type of assay to be used (see below), either the antigen or antibody is initially linked to an enzyme to give a conjugate which has both enzymatic and immunological reactivity. The assay is then carried out with either antigen or antibody immobilised on a solid surface. This immobilisation allows for simple washing steps that are necessary to separate reacted labelled reagent from unreacted reagent. Having incubated antigen with antibody, and having carried out appropriate washing steps to remove excess reagents, the enzyme activity remaining in the bound fraction is then quantitated by the addition of a non-chromatic substrate which is converted by the linked enzyme to a highly chromatic product.

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Further Reading

  • Clark B.R. and Engvall, E. (1980) Enzyme Linked Immunosorbent Assay (ELISA). Theoretical and Practical Aspects, in Maggio, E.T. (ed.) Enzyme Immunoassay, pp. 167–180 (CRC Press, Boca Raton, Florida)

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  • Engvall, E. (1980) Enzyme Immunoassay ELISA and EMIT. Methods in Enzymology, 70, pp. 419–439 (Academic Press, London)

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  • Proceedings of the meeting Recent Developments in ELISA and Other Solid Phase Immunoassays, held at Guy’s Hospital, London, 18–19 May 1985. Journal of Immunological Methods (1986) Vol. 87, No. 1

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  • Voller, A. (1980) Heterogeneous Enzyme-immunoassays and Their Applications, in Maggio, E.T. (ed.) Enzyme Immunoassay, pp. 181–195 (CRC Press, Boca Raton, Florida)

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  • Voller, A., Bidwell, D.E. and Bartlett, A. (1979) The Enzyme-linked Immunosorbent Assay (ELISA) (Dynatech Europe, Guernsey)

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  • Voller, A. and Bidwell, D.E. (1981) The Enzyme Linked Immunosorbent Assay (ELISA), Volume 2 (Microsystems Ltd., Guernsey)

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  • Voller, A., Bartlett, A. and Bidwell, D. (1981) Immunoassays for the Eighties (MTP Press, Lancaster)

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  • Voller, A. and Bidwell, D.E. (1982) The Enzyme Linked Immunosorbent Assay (ELISA). Med. Lab. World, March

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  • Wood, H.C. (1982) Practical Aspects of Heterogeneous Enzyme Immunoassays. Qualityline, Summer. A free brochure provided by Miles Research Products Division

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Authors
  1. John M. Walker
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Editors and Affiliations

  1. Division of Biological and Environmental Sciences, The Hatfield Polytechnic, PO Box 109, College Lane, Hatfield, Hertfordshire, AL10 9AB, England

    John M. Walker (Principal Lecturer in Biochemistry) (Principal Lecturer in Biochemistry)

  2. Department of Infectious Diseases, Section of Bacteriology, University of Utrecht, PO Box 80.171, 3508 TD, Utrecht, The Netherlands

    Wim Gaastra (Associate Professor of Microbiology) (Associate Professor of Microbiology)

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© 1987 John M. Walker and Wim Gaastra

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Walker, J.M. (1987). The Enzyme Linked Immunosorbent Assay (ELISA). In: Walker, J.M., Gaastra, W. (eds) Techniques in Molecular Biology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-9799-5_4

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  • DOI: https://doi.org/10.1007/978-1-4615-9799-5_4

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-0-7099-3673-2

  • Online ISBN: 978-1-4615-9799-5

  • eBook Packages: Springer Book Archive

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