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Anal Chem. 2010 Feb 15;82(4):1498-508. doi: 10.1021/ac902617t. Mass spectrometric-based stable isotopic 2-aminobenzoic acid glycan mapping for rapid glycan screening of biotherapeutics. Prien JM1, Prater BD, Qin Q, Cockrill SL. Abstract
Fast, sensitive, robust methods for "high-level" glycan screening are necessary during various stages of a biotherapeutic product's lifecycle, including clone selection, process changes, and quality control for lot release testing. Traditional glycan screening involves chromatographic or electrophoretic separation-based methods, and, although reproducible, these methods can be time-consuming. Even ultrahigh-performance chromatographic and microfluidic integrated LC/MS systems, which work on the tens of minute time scale, become lengthy when hundreds of samples are to be analyzed. Comparatively, a direct infusion mass spectrometry (MS)-based glycan screening method acquires data on a millisecond time scale, exhibits exquisite sensitivity and reproducibility, and is amenable to automated peak annotation. In addition, characterization of glycan species via sequential mass spectrometry can be performed simultaneously. Here, we demonstrate a quantitative high-throughput MS-based mapping approach using stable isotope 2-aminobenzoic acid (2-AA) for rapid "high-level" glycan screening.
Nanospray Mass Spectrometry (NSI-MS). Negative- and positive-mode mass spectrometric data of directly infused sample mixtures was obtained on a linear ion trap mass spectrometer (Thermo LTQ XL, San Jose, CA) equipped with a TriVersa Nanomate-automated nanospray ion source (Advion, Ithaca, NY).