Development and optimization of cytokine ELISAs using commercial antibody pairs

The measurement of cytokines in plasma and other fluids often requires the use of an enzyme-linked immunosorbant assay (ELISA). In the research environment, a valuable assay is one that yields reliable results in the shortest amount of time for the least cost. To achieve this goal, a protocol has be...

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Bibliographic Details
Published in:Journal of immunological methods 2001-09, Vol.255 (1), p.149-157
Main Authors: Nemzek, Jean A, Siddiqui, Javed, Remick, Daniel G
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Antibodies
Antibodies - economics
Biological and medical sciences
Buffer
Cytokine
Cytokines - analysis
ELISA
Enzyme-Linked Immunosorbent Assay - economics
Enzyme-Linked Immunosorbent Assay - methods
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Indicators and Reagents - economics
Interleukin-6 - analysis
Microbiology
Molecular immunology
Protein hormones. Growth factors. Cytokines
Proteins
Techniques
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Summary:The measurement of cytokines in plasma and other fluids often requires the use of an enzyme-linked immunosorbant assay (ELISA). In the research environment, a valuable assay is one that yields reliable results in the shortest amount of time for the least cost. To achieve this goal, a protocol has been outlined to develop sandwich ELISAs for cytokines using commercial antibodies. These guidelines for ELISA development include selecting antibody concentrations, choosing an appropriate buffer, reducing plasma interference and evaluating the optimal length for incubation periods. In addition, the protocol for a rapid IL-6 ELISA is presented. This ELISA allows measurement of IL-6 in a reduced amount of time by raising the concentration of antibodies used and increasing the temperature for incubation. By following the guidelines presented, cost-effective, cytokine ELISAs can be developed that yield low background, detect a wide range of concentrations, and are suitable for use in the research setting.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(01)00419-7