Selection of Antibodies with Tailored Properties by Application of High-Throughput Multiparameter Fluorescence-Activated Cell Sorting of Yeast-Displayed Immune Libraries

Christian Schröter,Stefan Becker,Harald Kolmar,Jan Beck,Stefan Zielonka,Björn Hock,Lars Toleikis,Simon Krah,Achim Doerner,Laura Rhiel,Ralf Günther

doi:10.1007/s12033-018-0109-0

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.

Highlights

Monoclonal antibodies as well as other proteins with antibody-like binding characteristics are among the most successful entities in the biopharmaceutical sector [1]
yeast surface display (YSD) libraries were constructed based on antibody repertoires from immunized OmniRatsTM [10, 27], which are transgenic for the expression of human antibodies
With respect to Monoclonal antibodies (mAbs) engineering, YSD proved to be a valid tool for affinity maturation [19, 20, 23] as well as for the direct isolation of highaffinity mAbs based on immune repertoires of rodents [19, 23]

Summary

Introduction

Monoclonal antibodies (mAbs) as well as other proteins with antibody-like binding characteristics are among the most successful entities in the biopharmaceutical sector [1]. Sources of antibody gene pools can be synthetic libraries or derived from B cell-receptor (BCR) gene repertoires after immunization requiring either wild-type or transgenic animals [3]. Libraries from immunized rodents commonly yield mAbs with high affinity and target specificity [8]. This is due to a strong immune response as a result of repeated challenge with a certain antigen. With the advent of transgenic animals harboring human immunoglobulin loci, fully human antibodies are raised by immunization, obviating the need for time-consuming humanization of mAbs isolated from wild-type animals [9, 10]

Methods

Results

Conclusion