NanoLC/MS Using 10 μm i.d. RP- and HILIC-PLOT Columns for Ultratrace Analysis

  报告时间20141212日(星期五)上午930 

  报告地点:生物楼406房间 

  报告人Quanzhou LuoBarnett Institute, Northeastern University, Boston, MA 02115 

    

  报告人简介 

  PROFESSTIONAL EXPERIENCE 

  2008–Present Amgen Inc. Thousand Oaks, CA  Senior Scientist  

  ·Managed team of scientist and associate scientist in areas of analytical chemistry and  mass spectrometry to support process and product development, methods development/qualification/transfer; product characterization 

  ·Experienced process development analytical science team leader for protein therapeutics, including antibody drug conjugate, monoclonal antibody and Fc fusion protein  

  ·Lead analytical development for biosimilar project.  

  ·Lead cross-function team on platform method development 

  ·Lead high end mass spectrometry team to support process development, including cell line development, reference standard characterization, and comparability study for regulatory filing 

  ·Co-lead mass spectrometry in QC initiative in Amgen 

  ·Authoring CMC and regulatory filings 

   2006–2008 Northeastern University, Dr. Karger Group, Boston, MA  Senior Scientist  

  ·Established a hydrophilic interaction chromatography (HILIC) LC/MS platform for ultratrace glycan characterization and structure analysis using MSn 

  ·Identified multiple proteins involved in major metabolic and regulatory pathways from laser capture microdissection (LCM) sample of 15, 000 lymphopatic breast cancer cells 

  2003–2006 Pacific Northwest National Lab, Dr. Smith Group, Richland, WA  Postdoc       

  ·Developed a nanoLC/MS platform for proteomic analysis using 20-μm-i.d. silica-based monolithic column 

  ·Co-invented a novel method for making monolithic electrospray ionization emitters  

  ·Developed an immobilized metal affinity chromatography (IMAC)/RPLC/MS/MS platform for sensitive phosphoproteome analysis 

  2002–2003 UC-Berkeley, Dr. Frechét/ Dr. Svec Group    Berkeley, CA    Postdoc  

  ·Co-invented novel matrix for surface-enhanced laser desorption/ionization time-of-flight mass spectrometry of small molecules 

  ·Developed monolithic valves for microfluidic chips based on thermoresponsive polymer 

  EDUCATION 

  Ph.D.  July 2002 Analytical chemistry, National Chromatographic R&A Center, Dalian  

                    Institute of Chemical Physics, Chinese Academy of Sciences, China 

  M.S.   July 1999 Analytical chemistry, Northwest University, China 

  B.S.   July 1997 Chemistry, Northwest University, China 

    

AWARDS & HIGHLIGHTS

  Amgen Acclaim Award–Product quality attributes assessment, 10/2013 

  P&PD Award–Pipeline Project Execution for AMG 145, Amgen Inc., 2012 

  P&PD Award–Innovation Award “Quantification of Posttranslational Modifications in Recombinant Proteins Using Stable Isotope Labeled Internal Standard and Mass Spectrometry”, Amgen Inc., 2011 

  P&PD Award–Cross Functional Collaboration “Immunogenicity of protein aggregates”, Amgen Inc., 2010 

  Research reported on American Laboratory News, a magazine published by International Scientific Communications, Inc., April, 2008 

  The article, "Ultratrace LC/MS Proteomic Analysis Using 10- μm-i.d. Porous Layer Open Tubular Poly(styrene-divinylbenzene) Capillary Columns", published in Analytical Chemistry is being featured on the ACS Publications website as one of the  Most-Cited Articles published in 2007  

  Research reported as Currents on Journal of Proteome Research, a scientific journal published by the American Chemical Society, February, 2007.

  Research reported as COVER STORY on Chemical & Engineering News, a magazine published by the American Chemical Society, December, 2006.  

  Gustel and Ernst Giessen Memorial Award in Advanced Research, Barnett Institute, 2006/2007 

  President Excellence Scholarship, Chinese Academy of Sciences, 2002 

  Outstanding Graduate Scholarship, Shaanxi Province, 1999 

  Outstanding Student Scholarship, Northwest University, Xi’an, 1996, 1995, 1994 

  报告简介 

  NanoESI-MS at low nL/min flow rate provides significant advantages over conventional ESI-MS, including improved sensitivity, increased ionization efficiency, reduced charge competition and ion suppression.  Therefore, new LC/MS approaches using ultranarrow-bore LC columns is highly useful for the analysis of limited sample amounts, e.g. samples containing several thousand specific cells collected by laser capture microdissection.  Recently, we developed a robust protocol for production of 10 ?m i.d. porous layer open tubular polystyrene-divinylbenzene columns operating at flow rates of 20 nL/min.1  To improve the ability to handle sample of few microliters volume and the resolving power of PLOT column, we developed on-line 1D and 2D PLOT/MS platforms.2,3  The new platform was used for comprehensive characterization of the proteome of SiHa cervical cancer cell line with the injection amount equivalent to 1200 cells.  In total, over 5000 peptides covering close to 2000 unique proteins were identified, including a number of proteins relevant to cancer, e.g. MAP kinases.   

  Several challenges exist for the LC/ESI/MS analysis of polar compounds, such as glycans, including (1) new chromatographic approaches to resolve such polar analytes effectively and (2) minimization of the ion suppression of these compounds during ESI.  To address these issues, we developed a new approach for the preparation of hydrophilic interaction chromatographic (HILIC) PLOT columns.  The amine-HILIC PLOT columns are characterized by high resolving power for glycans at a flow rate of 20 nL/min.  When coupled on-line with ESI-MS, the columns demonstrated good sensitivity for the analysis of polar compounds, which are usually difficult to be detected due to ion suppression.  The use of such columns will be described in the presentation.  

  报告联系人:1809组 王璐(9620 

版权所有 © 中国科学院大连化学物理研究所 本站内容如涉及知识产权问题请联系我们 备案号:辽ICP备05000861号-1 辽公网安备21020402000367号