Saccharomyces cerevisiae strain:EBY100 Raw sequence reads

To ensure a fast and effective immune response, B-cells produce antibodies that bind specifically to foreign antigens. Despite the central role that antibodies play in the adaptive immune system and in biotechnology, much remains unknown about the quantitative re- lationship between an antibody's peptide sequence and its antigen binding affinity. Recently, a variety of deep mutational scanning tech- nologies have been described for assaying protein sequence-function relationships. These approaches, however, are largely unable to de- convolve the sequence-dependence of binding affinity from confound- ing effects on expression and protein stability. Here we describe a new experimental approach, called Tite-Seq, that is capable of mea- suring binding titration curves for thousands of different proteins in parallel. These measurements eliminate the confounding effects of expression and stability, enabling the direct observation of how pro- tein sequence affects binding. We demonstrate this method through measurements of how the CDR1H and CDR3H regions of a well- studied scFv antibody affect binding to fluorescein. Tite-Seq fills a large gap in our ability to measure critical aspects of the adaptive immune system, and can be readily used for studying the sequence dependence of binding affinity in many other protein systems.