Precise and Efficient Antibody Epitope Determination Through Library Design, Yeast Display and Next-Generation Sequencing

Van Blarcom T, et al., 427(6 Pt B):1513-34, J Mol Biol, 2015

This article describes a methodology for epitope mapping of a panel of antibodies over a course of several weeks. It relies on rational library design, yeast display, and the next-gen DNA sequencing. The model system used in this article includes several neutralizing antibodies that bind to alpha toxin from Staphylococcus aureus. The affinity estimate is expected to serve as a predictor of antibody potency and hence the best clinical candidates. The epitope binning experiments performed using an Octet RED384 instrument equipped with Streptavidin biosensors allowed capture of biotinylated antibodies. Then the alpha toxin was bound to these captured antibodies. The pair wise binding was assessed in a classical sandwich orientation using the unbiotinylated antibodies.

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