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Amplification Systems for Enzyme Immunoassay

  • Jean-Luc Guesdon

Abstract

Since its development (Avrameas and Guilbert, 1971a, 1971b; Engvall and Perlmann, 1971; Van Weemen and Schuurs, 1971) the enzyme immunoassay has been applied extensively for diagnosing bacterial, viral and parasitic diseases. The enzyme immunoassay method has proved to be a powerful tool in many areas and has been the focus of several reviews, symposia and workshops (Feldmann et al., 1976; Engvall and Pesce, 1978; Malvano, 1980; Guesdon and Avrameas, 1981; Ishikawa et al., 1981; Avrameas et al., 1983; Talwar, 1983). In order to broaden the application of the enzyme immunoassay, it is important to improve existing technology; indeed, highly sensitive assays are required in fundamental research as well as in medicine. The enzyme immunoassay, involving antigen, hapten or antibody labelled with an enzyme, combines the specific recognition properties of antibodies with the high sensitivity characteristic of enzyme based analytical techniques. The physicochemical properties and the concentration of the antibodies and the enzyme directly affect the assay’s sensitivity. More precisely, the best sensitivity is obtained with antibodies having a high affinity constant. Whereas the affinity constant cannot be increased beyond its upper limit, the sensitivity of the competitive immunoassay can be improved by decreasing the concentration of antibody used. Indeed, the effect of decreasing the antibody concentration is to shift the usable range to a lower concentration of antigen. As the antibody concentration decreases, however, the dose-response curve flattens and higher sensitivity is achieved at the expense of discrimination: consequently, the assay will be less accurate. Thus, using an enzyme-antibody conjugate with high immunoreactivity and enzyme activity, and amplifying the specific signal so that a low antibody concentration may be used will improve both the sensitivity and the accuracy of the enzyme immunoassay.

Keywords

Enzyme Immunoassay Glucose Oxidase Fluorogenic Substrate Biospecific Interaction Rabbit Immune Serum 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Jean-Luc Guesdon
    • 1
  1. 1.Laboratoire des Sondes FroidesInstitut PasteurParisFrance

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