Combining phage display with de novo protein sequencing for reverse engineering of monoclonal antibodies.

ABSTRACT: The enormous diversity created by gene recombination and somatic hypermutation makes de novo protein sequencing of monoclonal antibodies a uniquely challenging problem. Modern mass spectrometry-based sequencing will rarely, if ever, provide a single unambiguous sequence for the variable domains. A more likely outcome is computation of an ensemble of highly similar sequences that can satisfy the experimental data. This outcome can result in the need for empirical testing of many candidate sequences, sometimes iteratively, to identity one which can replicate the activity of the parental antibody. Here we describe an improved approach to antibody protein sequencing by using phage display technology to generate a combinatorial library of sequences that satisfy the mass spectrometry data, and s

SUBMITTER: Rickert KW 

PROVIDER: S-EPMC4966834 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature