Part of the Methods in Molecular Biology™ book series (MIMB, volume 1)
Enzyme-Linked Immunosorbant Assay (ELISA)
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.
KeywordsH2O2 Albumin EDTA Ethylene Glycol H2SO4
© Humana Press 1984