Enzyme-linked immunoassay (ELISA) for connective tissue components

S. I. Rennard, R. Berg, G. R. Martin, J. M. Foidart, P. Gehron Robey

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Abstract

Enzyme-linked immunoadsorbant assays have been developed for types I, II, III, and IV collagen and for laminin and fibronectin. These assays offer a specific, sensitive, and convenient method for the measurement of various connective tissue components either separately or simultaneously. Plastic microtiter wells are coated with purified antigen, then antibodies to the antigen are allowed to bind to the insolubilized antigen in each well. The amount of bound antibody is determined by incubation with a second antibody which is covalently linked to alkaline phosphatase or horseradish peroxidase. The amount of bound enzyme is assayed after adding an appropriate substrate. The level of protein in a sample is estimated from its ability to inhibit the binding of the first antibody to the antigen-coated well. Specificity of the assay depends on the purity of the adsorbed antigen and allows for highly specific tests. Under optimum conditions the sensitivity of the assay is determined by the KB of the antibody and allows 10-20 ng of a specific type of collagen or laminin and 1 ng fibronectin to be detected.

Original languageEnglish (US)
Pages (from-to)205-214
Number of pages10
JournalAnalytical Biochemistry
Volume104
Issue number1
DOIs
Publication statusPublished - May 1 1980

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Rennard, S. I., Berg, R., Martin, G. R., Foidart, J. M., & Robey, P. G. (1980). Enzyme-linked immunoassay (ELISA) for connective tissue components. Analytical Biochemistry, 104(1), 205-214. https://doi.org/10.1016/0003-2697(80)90300-0