Abstract
In general, immunological methods are not very well suited for a quantitative determination of the antigen to be studied. The ELISA technique, however, can be used for a quantitative or at least semiquantitative determination of the concentration of a certain antigen. The method was first introduced by Engvall and Perlmann (1). The principle of ELISA (see Fig. 1), also called the double antibody sandwich technique, is the following: Antibodies against the antigen to be measured are adsorbed to a solid support, in most cases a polystyrene microtiter plate. After coating the support with antibody and washing, the antigen is added and will bind to the adsorbed antibodies. Next, a conjugate that will also bind to the antigen is added. Conjugates are antibody molecules to which an enzyme is covalently bound.
The main steps in a (noncompetitive) ELISA test. (1) The antibody to the antigen being quantitated is adsorbed onto a solid phase, usually polystyrene. (2) The sample containing the antigen being measured is then added. (3) Following the incubation and washing steps, a second enzyme-labeled antibody is then added. After further incubation and washing steps, enzyme substrate is added. (A substrate is chosen that will give a colored product). The amount of color produced is therefore proportional to the amount of antigen bound to the original antibody.
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References
Engvall, E., and Perlmann, P. (1971) Immunochemistry 8, 871–874.
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© 1984 Humana Press
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Gaastra, W. (1984). Enzyme-Linked Immunosorbant Assay (ELISA). In: Walker, J.M. (eds) Proteins. Methods in Molecular Biology™, vol 1. Humana Press. https://doi.org/10.1385/0-89603-062-8:349
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DOI: https://doi.org/10.1385/0-89603-062-8:349
Publisher Name: Humana Press
Print ISBN: 978-0-89603-062-6
Online ISBN: 978-1-59259-488-7
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