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

Time: Dec. 12th 2014 9:30 AM

Location: Biotechnology Building, Room 406

Lecturer: Quanzhou Luo(Barnett Institute, Northeastern University, Boston, MA 02115) 

 

Introduction:

PROFESSTIONAL EXPERIENCE

 

2008 – Present Amgen Inc. Thousand Oaks, CASeniorScientist

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

·Developed a nanoLC/MS platform for proteomic analysisusing 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 GroupBerkeley, CAPostdoc

·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

 

 

Abstract:

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.

Contact: Droup 1809 Wang Lu (9620)

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