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Enzyme-Linked Immunosorbent Assay

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.
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Updated: Jul 3, 2026

Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting

Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting

Published on: December 12, 2011

A rapid, simple immunofluorometric assay: development and characterization.

H Park¹, D I Wang, M L Yarmush

¹Department of Applied Biological Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Biotechnology and Bioengineering

|June 20, 1992

Summary

This summary is machine-generated.

A novel immunofluorometric assay uses small beads and measures unbound antibodies for rapid protein detection. This assay offers wide-ranging antigen quantification without sample dilution, improving efficiency in diagnostics.

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Area of Science:

Biochemistry
Immunology
Analytical Chemistry

Background:

Traditional immunoassays often involve multiple steps and longer incubation times.
Accurate and rapid quantification of proteins is crucial for diagnostics and research.

Purpose of the Study:

To develop and validate a novel, rapid, two-site immunofluorometric assay.
To demonstrate the assay's applicability for quantifying proteins over a wide concentration range without dilution.

Main Methods:

Utilized small beads (0.5 microm) as a solid support in a one-step incubation and one-step separation assay.
Employed a two-site immunofluorometric approach measuring the unbound fraction of labeled antibody.
Developed specific assays for human IgG (polyclonal antibodies) and bovine serum albumin (BSA) (monoclonal antibodies).

Main Results:

The human IgG assay detected concentrations from 0.2 to 40 microg/mL with a 30-minute incubation.
The BSA assay detected concentrations from 0.2 to 14 microg/mL with a rapid 2-minute incubation.
Demonstrated low variability for BSA measurement (interassay 1.9%, intra-assay 2.3%).

Conclusions:

The novel assay is rapid, reproducible, and suitable for quantifying proteins over a broad dynamic range.
The assay's principle is adaptable for measuring various proteins, offering a versatile diagnostic tool.