120th General Meeting of the KCS

Type Oral Presentation
Area Oral Presentation of Young Analytical Chemists I
Room No. Room C311+C312
Time THU 10:05-:
Code ANAL1.O-23
Subject On-line proteolysis and glycopeptide enrichment using dual micro-scale porous polymer membrane enzyme reactor (μPPMER) and nanoflow liquid chromatography-tandem mass spectrometry
Authors JoonSeon Yang, Juan Qiao1, Liping Zhao2, Li Qi2,*, Myeong Hee Moon*
Department of Chemistry, Yonsei University, Korea
1Beijing National Laboratory for Molecular Sciences; Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Acedemy of Sciences, Chile
2Beijing National Laboratory for Molecular Sciences; Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Acedemy of Sciences, China
Abstract N-glycosylation is a type of glycosylation that occurs when glycans are attached to the nitrogen atom of asparagine (asn, N). It is important to analyze N-glycoproteins in biological samples as they play essential roles in protein folding or signaling process. However, analysis of glycoproteins from glycopeptides by shotgun analysis can be time-consuming and sample loss is inevitable during pre-treatments. Therefore, to overcome such problems, dual micro-scale thermo-sensitive porous polymer membrane enzyme reactors (μPPMER) has been utilized for on-line proteolysis, followed by enrichment of glycopeptides prior to nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS). A thermo-responsive porous polymer membrane (T-PPM) was synthesized by coating PS-co-MAn-NIPAM polymer on bare nylon membrane and its characterization was conducted by SEM. Trypsin and lectin were immobilized on the coated membrane for proteolysis and glycopeptide capture, respectively, and each of membrane was inserted in μPPMER module. Dual μPPMER modules were connected to nLC-ESI-MS/MS system for on-line analysis and efficiency in glycopeptides enrichment selectivity under various temperatures were evaluated. In addition, glycoproteins from human plasma and urine samples were analyzed using on-line dual PPMER module in the most optimized condition for glycopeptide enrichment.
E-mail js0610@yonsei.ac.kr