Abstract
Understanding antibody specificity and defining response profiles to antigens continue to be essential to both vaccine research and therapeutic antibody development. Peptide scanning assays enable map** of continuous epitopes in order to delineate antibody–antigen interactions beyond traditional immunoassay formats. We have developed a relatively low-cost method to generate peptide microarray slides for antibody binding studies that allow for interrogation of up to 1536 overlap** peptides derived from the target antigens on a single microslide. Using an IntavisAG MultiPep RS peptide synthesizer and a Digilab MicroGrid II 600 microarray printer robot, each peptide is tagged with a polyethylene glycol aminooxy terminus to improve peptide solubility, orientation, and conjugation efficiency to the slide surface. Interrogation of the surface can then be performed using polyclonal immune sera or monoclonal antibodies, and sensitive detection using an InnoScan 1100 AL scanner with fluorescent-conjugated secondary reagents maximizes conservation of reagents.
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Acknowledgements
This work was partly supported by NIH awards AI158193 and AI168917 to ML, and a David C. Fairchild Endowed Fellowship, Skaggs Graduate School of Chemical and Biological Sciences, The Scripps Research Institute, to KN.
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Nagy, K., McBride, R., Head, S.R., Ordoukhanian, P., Law, M. (2023). Low-Cost Peptide Microarrays for Map** Continuous Antibody Epitopes. In: Cretich, M., Gori, A. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 2578. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2732-7_6
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DOI: https://doi.org/10.1007/978-1-0716-2732-7_6
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