RT Journal Article
SR Electronic
T1 Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 036335
DO 10.1101/036335
A1 Rhys M. Adams
A1 Thierry Mora
A1 Aleksandra M. Walczak
A1 Justin B. Kinney
YR 2016
UL http://biorxiv.org/content/early/2016/11/15/036335.abstract
AB 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 TiteSeq, 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.