Next generation VAMS®–Trypsin immobilization for instant proteolysis in bottom-up protein determination

نویسندگان

چکیده

The purpose of this study was to develop a smart proteolysis sampler based on commercially available VAMS® tips employing Volumetric Absorptive Microsampling. In proof-of-concept paper we show that covalently bound trypsin VAMS exhibits sufficient activity for proteomic purposes without altering physical conditions. Alkaline pH needed the covalent binding and at least 50 μg required obtain maximal activity. Prolonged digestion times (up overnight) sample containing 7 model proteins yielded higher peak intensities signature peptides which are beneficial quantitative analysis. In-solution digestions three (carbonic anhydrase 2, catalase BSA) their whereas other four (cytochrome C, myoglobin, β-lactoglobulin, phosphorylase B) showed from performed modified VAMS. comparison with conventional in-solution digestions, approximately half were identified diluted serum using For shotgun proteomics, samples collected followed by immediate drying similar peptide- (942 vs 875) protein (96 99) identifications compared those overnight digestions. This result shows potential

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ژورنال

عنوان ژورنال: Advances in Sample Preparation

سال: 2022

ISSN: ['2772-5820']

DOI: https://doi.org/10.1016/j.sampre.2022.100027