A Liquid Chromatography/Mass Spectrometry Method for Screening Disulfide Tethering Fragments.

ABSTRACT: We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a percent bound value for each compound and identifies false positiv

SUBMITTER: Hallenbeck KK 

PROVIDER: S-EPMC6609441 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature