Generation and validation of recombinant antibodies to study human aminoacyl-tRNA synthetases
Charlotta Preger, Edvard Wigren, Elena Ossipova, Carolyn Marks, Johan Lengqvist, Camilla Hofström, Oskar Andersson, Per‐Johan Jakobsson, S. Gräslund, Helena Persson
Abstract
and used as antigens in phage display selections using a synthetic human single-chain fragment variable library. All targets yielded large sets of antibody candidates that were validated through a panel of binding assays against the purified antigen. Furthermore, the top-performing binders were tested in immunoprecipitation followed by MS for their ability to capture the endogenous protein from mammalian cell lysates. For antibodies targeting individual members of the multi-tRNA synthetase complex, we were able to detect all members of the complex, co-immunoprecipitating with the target, in several cell types. The functionality of a subset of binders for each target was also confirmed using immunofluorescence. The sequences of these proteins have been deposited in publicly available databases and repositories. We anticipate that this open source resource, in the form of high-quality recombinant proteins and antibodies, will accelerate and empower future research of the role of aaRSs in health and disease.