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Clinical Chemistry 27: 1602-1608, 1981;
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Clinical Chemistry, Vol 27, 1602-1608, Copyright © 1981 by American Association for Clinical Chemistry

Enzyme-enhancement immunoassay: a homogeneous assay for polyvalent ligands and antibodies

I Gibbons, TM Hanlon, CN Skold, ME Russell and EF Ullman

A homogeneous enzyme immunoassay for proteins has been developed that avoids the need for a labeled antigen. The technique involves antibody labeled with beta-galactosidase (EC 3.2.1.23), succinylated antibody, and a macromolecular o-nitrophenyl-beta-galactoside substrate. The enzyme-labeled antibody and the succinylated antibody form an immune complex in the presence of sample antigen. An enzyme within this negatively charged microenvironment produces a product that forms a second light-scattering phase, whereas the product produced by free enzyme remains soluble. Thus the antigen modulates the rate of increase in light scattering. The technique has been applied to assays for human immunoglobulin G and C-reactive protein as well as for specific antibodies.




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Copyright © 1981 by the American Association for Clinical Chemistry.