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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.
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.

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Related Experiment Video

Updated: Jun 18, 2026

Measuring the 50% Haemolytic Complement (CH50) Activity of Serum

Measuring the 50% Haemolytic Complement (CH₅₀) Activity of Serum

Published on: March 29, 2010

A standardized method for quantitating the complement-mediated immune complex solubilizing capacity of human serum.

G Baatrup, I Peterson, S E Svehag

Journal of Immunological Methods

|May 13, 1983

Summary

A new radioassay standardizes measurement of complement-mediated immune complex solubilizing capacity (CMSC). This assay shows reduced CMSC in SLE sera, offering insights into immune complex disorders.

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Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

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Published on: December 14, 2016

Related Experiment Videos

Last Updated: Jun 18, 2026

Measuring the 50% Haemolytic Complement (CH50) Activity of Serum

Measuring the 50% Haemolytic Complement (CH₅₀) Activity of Serum

Published on: March 29, 2010

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

Published on: January 29, 2014

Optimization of a Quantitative Micro-neutralization Assay

Optimization of a Quantitative Micro-neutralization Assay

Published on: December 14, 2016

Area of Science:

Immunology
Biochemistry
Assay Development

Background:

Immune complexes (ICs) play a role in various autoimmune diseases.
Assessing the capacity of complement (C) to solubilize ICs is crucial for understanding disease pathogenesis.
Standardized assays are needed for reliable measurement of complement-mediated solubilization.

Purpose of the Study:

To describe a standardized radioassay for measuring complement-mediated immune complex solubilizing capacity (CMSC).
To evaluate the initial kinetics of the solubilization (IKS) reaction.
To optimize and validate the assay for clinical and research applications.

Main Methods:

Developed a radioassay using 125I-BSA-anti-BSA immune complexes and diluted serum.
Measured percentage C-mediated solubilization (CMS) by quantifying radioactivity distribution post-centrifugation.
Investigated assay dependency on serum dilution, buffer, IC amount, storage, and centrifugation.

Main Results:

Established a standardized assay with a coefficient of variation (CVt) of 0.10-0.17.
Demonstrated that treatments inactivating or interfering with complement significantly reduce CMSC.
Observed reduced IKS and CMSC in systemic lupus erythematosus (SLE) sera.
Found normal IKS and only marginal CMSC reduction in rheumatoid arthritis sera.

Conclusions:

The developed radioassay provides a reliable method for quantifying CMSC and IKS.
Assay results indicate impaired complement-mediated immune complex clearance in SLE.
Findings suggest potential diagnostic or prognostic value of CMSC and IKS measurements in autoimmune diseases.