Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves

Adams, R. M., Mora, T., Walczak, A. M., Kinney, J. B. (2016) Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves. Elife, 5. ISSN 2050-084X (Electronic)2050-084X (Linking)

URL: https://www.ncbi.nlm.nih.gov/pubmed/28035901
DOI: 10.7554/eLife.23156

Abstract

Despite the central role that antibodies play in the adaptive immune system and in biotechnology, much remains unknown about the quantitative relationship between an antibody's amino acid sequence and its antigen binding affinity. Here we describe a new experimental approach, called Tite-Seq, that is capable of measuring binding titration curves and corresponding affinities for thousands of variant antibodies in parallel. The measurement of titration curves eliminates the confounding effects of antibody expression and stability that arise in standard deep mutational scanning assays. We demonstrate Tite-Seq on the CDR1H and CDR3H regions of a well-studied scFv antibody. Our data shed light on the structural basis for antigen binding affinity and suggests a role for secondary CDR loops in establishing antibody stability. Tite-Seq fills a large gap in the ability to measure critical aspects of the adaptive immune system, and can be readily used for studying sequence-affinity landscapes in other protein systems.

Item Type: Paper
Uncontrolled Keywords: S. cerevisiae biophysics structural biology
Subjects: structural biology
organism description > yeast
CSHL Authors:
Communities: CSHL labs > Kinney lab
Depositing User: Matt Covey
Date Deposited: 04 Jan 2017 19:20
Last Modified: 07 Sep 2017 14:41
Related URLs:
Dataset ID:
URI: http://repository.cshl.edu/id/eprint/33955

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