겨울 심포지움
2018겨울초록
발표자 및 발표 내용
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접수자
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Protein N-glycosylation, one of post-translational momdifications (PTMs), is a process that glycans attach covalently to nitrogen atom in asparagine (asn, N) amino acid. Since several glycosylated proteins are known as biomarkers of disease such as cancers, it is important to analyze glycoproteins in biological samples. In this study, dual micro-scale thermo-sensitive porous polymer membrane reactor (TPPMR) was developed for on-line proteolysis and glycopeptide enrichment prior to nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS).
PS-MAn-NIPAM (PNPAm) is thermo-sensitive porous polymer membrane which is synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization and is coated on nylon membrane by breath figure method. Trypsin and lectin mixtures (ConA and WGA) can be immobilized on the coated membrane for proteolysis and glycopeptide enrichment, respectively, and each of membrane is inserted in TPPMR module. Dual TPPMR modules are directly connected to nLC-ESI-MS/MS system and proteolysis efficiency in proteolysis and glycopeptides enrichment were evaluated by varying temperature. The developed method demonstrates that on-line proteolysis and glycopeptide enrichment can be performed with only about 1.5 μL human plasma sample with the successful identification of 155 glycoproteins.
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