Solid-phase immunoassays

Michael A Lynes1

  • 1University of Connecticut, Storrs, CT, USA.

Solid-phase quantitative immunoassays are some of the most commonly used diagnostic tests for both soluble antigen composition and the assessment of cellular functions. These immunoassays (e.g., ELISA, ELISPOT) provide a highly sensitive means to measure the presence of antigen in defined and homogeneous samples such as purified proteins in buffer, as well as in undefined heterogenous biological samples such as cell lysates, tissue culture supernatants, blood, and other clinical samples. The sensitivity of these assays can enable (under optimal conditions) detection of protein concentrations in the picogram range. A recent modification of the basic ELISA immunoassay takes advantage of the phenomenon of grating-coupled surface plasmon resonance (GCSPR) to provide a label-free real-time variant of this solid-phase immunoassay. Using GCSPR, similar assessments of antigen-antibody interactions can be done with smaller sample sizes and in a microarray format that enables the simultaneous measurement of large numbers of antibody/antigen interactions on the same sensor chip. These measurements allow for a highly refined and sensitive determination of the effects that toxins can have on biological systems, and they can be applied to a variety of immune and non-immune protein, cell, and tissue evaluations.

Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.