Selection of Antibodies with Tailored Properties by Application of High-Throughput Multiparameter Fluorescence-Activated Cell Sorting of Yeast-Displayed Immune Libraries |
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Authors: | Christian Schröter Jan Beck Simon Krah Stefan Zielonka Achim Doerner Laura Rhiel Ralf Günther Lars Toleikis Harald Kolmar Björn Hock Stefan Becker |
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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 |
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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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