T.O.P. (Team of pharmacoproteomics research)

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1 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. T.O.P. (Team of pharmacoproteomics research)

2 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.) 이지혜 (연구원)

3 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

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

5 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

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

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

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

9 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.

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

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

12 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.) 이지혜 (연구원)