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Selection of Antibodies with Tailored Properties by Application of High-Throughput Multiparameter Fluorescence-Activated Cell Sorting of Yeast-Displayed Immune Libraries
Authors:Christian Schröter  Jan Beck  Simon Krah  Stefan Zielonka  Achim Doerner  Laura Rhiel  Ralf Günther  Lars Toleikis  Harald Kolmar  Björn Hock  Stefan Becker
Affiliation:1.Antibody Drug Conjugates and Targeted NBE Therapeutics, Merck KGaA,Darmstadt,Germany;2.Institute for Organic Chemistry and Biochemistry,Technische Universit?t Darmstadt,Darmstadt,Germany;3.Protein Engineering and Antibody Technologies, Merck KGaA,Darmstadt,Germany
Abstract:In this study, we present a multiparameter screening procedure for the identification of target-specific antibodies with prescribed properties. Based on B cell receptor gene repertoires from transgenic rats, yeast surface display libraries were generated, and high-affinity human antibodies were readily isolated. We demonstrate that specific desirable features, i.e., species’ cross-reactivity and a broad epitope coverage can be integrated into the screening procedure using high-throughput fluorescence-activated cell sorting. We show that the applied screening stringencies translate directly into binding properties of isolated human antibody variants.
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