Mass Spectrometric-Based Stable Isotopic 2-Aminobenzoic Acid Glycan Mapping for Rapid Glycan Screening of Biotherapeutics

摘要

Fast, sensitive, robust methods for “high-level” glycannscreening are necessary during various stages of a biotherapeuticnproduct’s lifecycle, including clone selection,nprocess changes, and quality control for lot release testing.nTraditional glycan screening involves chromatographic ornelectrophoretic separation-based methods, and, althoughnreproducible, these methods can be time-consuming.nEven ultrahigh-performance chromatographic and microfluidicnintegrated LC/MS systems, which work on the tensnof minute time scale, become lengthy when hundreds ofnsamples are to be analyzed. Comparatively, a directninfusion mass spectrometry (MS)-based glycan screeningnmethod acquires data on a millisecond time scale, exhibitsnexquisite sensitivity and reproducibility, and is amenablento automated peak annotation. In addition, characterizationnof glycan species via sequential mass spectrometryncan be performed simultaneously. Here, we demonstratena quantitative high-throughput MS-based mapping approachnusing stable isotope 2-aminobenzoic acid (2-AA)nfor rapid “high-level” glycan screening.