An acid dissociation bridging ELISA for detection of antibodies directed against therapeutic proteins in the presence of antigen

Bridging Enzyme-Linked Immunosorbent Assay (ELISA) is commonly used in detection of antibodies directed against therapeutic proteins. Advantages of the bridging ELISA include the capability to detect antibodies regardless of their isotype or the species of origin. However, detection of antibodies ca...

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Bibliographic Details
Published in:Journal of immunological methods 2005-09, Vol.304 (1), p.189-195
Main Authors: Patton, Aaron, Mullenix, Michael C., Swanson, Steven J., Koren, Eugen
Format: Article
Language:English
Subjects:
Acid dissociation
Antibodies - analysis
Antibody assay
Antigen interference
Antigens - immunology
Biological and medical sciences
Calibration
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Humans
Molecular immunology
Proteins - immunology
Proteins - therapeutic use
Techniques
Therapeutic proteins
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Summary:Bridging Enzyme-Linked Immunosorbent Assay (ELISA) is commonly used in detection of antibodies directed against therapeutic proteins. Advantages of the bridging ELISA include the capability to detect antibodies regardless of their isotype or the species of origin. However, detection of antibodies can be difficult, if not impossible, in the presence of high levels of the antigen in the sample matrix. This protocol describes a bridging ELISA that uses a covalently coupled high density antigen surface combined with an acid dissociation step to allow for antibody detection in the presence of antigen in human serum.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2005.06.014