T.O.P. (Team of pharmacoproteomics research) TOP’s Service 질량분석 기반 단백질 정량 분석 서비스 Mass Spectrometry based- proteomics analysis Byung-Gyu Kim Ph.D. E-mail: Goldenlion@snu.ac.kr T.O.P. (Team of pharmacoproteomics research)

Summary of TOP’s Proteomics services 단백질분석시스템 분석내용 (항목) - 단백질질량분석기: Hybrid LC-MS (Resolution= up to 100,000) NanoHPLC system : EaZY nano LC II (Thermo Scientific) Mass analyzer: LTQ-Obitrap Velos ETD -데이터 처리 시스템: <Basic S/W> -Sorcerer 2/Scaffold 3 <Label-based quantification용 S/W> -Proteome discoverer 2.0 (CID, HCD, ETD 분석 가능) <Label-free Quantification용 S/W> -SIEVE ( label-free Quantification 용 S/W) -Progenesis (Multi group or Multi-Fractionation label-free Quantification 용 S/W) * MCP (Molecular and Cellular proteomics journal, IF=8.8) guide line에 맞는 data quality 제공 단백질 발현 비교분석 프로파일링 (Differentially Expressed Protein profiling) (B) 타겟 단백질 전사 후 수식부위 정밀분석 (Post-Translational Modification analysis) (C) 타깃 결합 단백질 프로파일링 (Interaction Proteome profiling) (D) 단백질 의약품 품질 정밀 분석 (Biosimillar Quality Control) (E) 약물 타깃 단백질 맵핑 (Drug target protein profiling) 문의 : 김병규 (Ph.D.) Goldenlion@snu.ac.kr 이지혜 (연구원) leejh@biocon.re.kr

TOP’s state-of-the-art Mass analyzer LTQ-Velos Obitrap ETD CID, PQD, MSA & ETD HCD Velos Obitrap ETD source √ Proteome Identification & quantification √ PTM site Mapping √ Drug target protein/site mapping √ Interaction partner mapping

TOP’s MS based-Proteome analysis Application ranges of TOP technologies nHPLC-LTQ-Obitrap-Velos-ETD analyzer Nat. biotech. 2010

TOP’s MS based-Proteome analysis 1st step Control Sample Cell Harvest Cell Lysates Sample 2 Sample 1 SDS-PAGE / Coomassie blue staining Protein 전처리

TOP’s MS based-Proteome analysis 2nd step Peptide 분리

TOP’s MS based-Proteome analysis 3rd step Mass 분석

TOP’s MS based-Proteome analysis 4th step Data 분석

TOP’s MS based-Proteome analysis Quantitative proteome analysis Metabolic Labeling WF Heavy peptides TMT0/2/6 ,mTRAQ, ICPL SILAC Label-free workflow Absolute Quan WF Isobaric Label WF Scaffold v3 Progenesis LC/MS Analysis SW Sample prep.

TOP’s MS based-Proteome analysis Analysis Eaxmple: Expression profile SILAC (Stable isotope labeled amino acid in cell culture) MS-analysis workflow Control Treated

TOP’s MS based-Proteome analysis Analysis Eaxmple: PTMs Phosphorylation easily identified by ETD CID/ETD Mode

Summary of TOP’s Proteomics services 단백질분석시스템 분석내용 (항목) - 단백질질량분석기: Hybrid LC-MS (Resolution= up to 100,000) NanoHPLC system : EaZY nano LC II (Thermo Scientific) Mass analyzer: LTQ-Obitrap Velos ETD -데이터 처리 시스템: <Basic S/W> -Sorcerer 2/Scaffold 3 <Label-based quantification용 S/W> -Proteome discoverer 2.0 (CID, HCD, ETD 분석 가능) <Label-free Quantification용 S/W> -SIEVE ( label-free Quantification 용 S/W) -Progenesis (Multi group or Multi-Fractionation label-free Quantification 용 S/W) * MCP (Molecular and Cellular proteomics journal, IF=8.8) guide line에 맞는 data quality 제공 단백질 발현 비교분석 프로파일링 (Differentially Expressed Protein profiling) (B) 타겟 단백질 전사 후 수식부위 정밀분석 (Post-Translational Modification analysis) (C) 타깃 결합 단백질 프로파일링 (Interaction Proteome profiling) (D) 단백질 의약품 품질 정밀 분석 (Biosimillar Quality Control) (E) 약물 타깃 단백질 맵핑 (Drug target protein profiling) 문의 : 김병규 (Ph.D.) Goldenlion@snu.ac.kr 이지혜 (연구원) leejh@biocon.re.kr