Abstract
Enzyme-linked immunosorbent assay (ELISA) is a commonly used method in analyzing biomolecular interactions. As a rapid, specific, and easy-to-operate method, ELISA has been used as a research tool as well as a widely adopted diagnostic method in clinical settings and for microbial testing in various industries. Inhibition ELISA is a one-site binding analysis method, which can monitor protein-protein interactions in solution as opposed to more commonly used sandwich ELISA in which the analyte capture step is required on a solid surface either through specific capture or through passive adsorption. Here, we introduce inhibition ELISA procedures, using a recombinant viral protein as an example, with emphasis on how inhibition ELISA could be used to probe subtle protein conformational changes in solution impacting protein–protein binding affinity. Inhibition ELISA is used to probe one binding site at a time for binding partners in solution with unrestricted conformation. The assay can be performed in a quantitative manner with a serially diluted analyte in solution for solution antigenicity or binding activity assessment.
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Acknowledgement
The work on chapter writing was enabled with the funding of Chinese Ministry of Science and Technology 863 Project (2012AA02A408) and National Science Foundation of China (81471934) to Q.Z.
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Weng, Z., Zhao, Q. (2015). Utilizing ELISA to Monitor Protein-Protein Interaction. In: Meyerkord, C., Fu, H. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 1278. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2425-7_21
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DOI: https://doi.org/10.1007/978-1-4939-2425-7_21
Publisher Name: Humana Press, New York, NY
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