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  English.dicp.cas.cn    Posted:2013-07-10
Glycomic Analysis by Glycoprotein Immobilization for Glycan Extraction and Liquid Chromatography on Microfluidic Chip

Time:2013-7-17  14:00 

Location:Academic Hall of Bio-tech building

Lecturer:Hui Zhang

Johns Hopkins University

Abstract:

Glycosylation is one of the most common protein modifications and profoundly regulates many biological processes. Aberrant glycosylation is reported to associate with diseases such as cancers, human immunodeficiency virus and immune disorders. It is considerably important to study protein glycosylation and the associated glycans for diagnostics and disease prognostics. Unlike other protein modifications, glycans attached to proteins are enormously complex. Therefore, the comprehensive analysis of glycans from biological or clinical samples is an unmet technical challenge. Development of the high-throughput method will facilitate the glycomics analysis. In this study we developed a novel method for the high-throughput analysis of N-glycans from glycoproteins using glycoprotein immobilization for glycan extraction (GIG) coupled with chipLC in an integrated microfluidic platform. The separated glycans were then analyzed by mass spectrometry. Briefly, proteins were first immobilized on a solid support. Glycans on immobilized glycoproteins were modified on solid phase to increase the detection and structure analysis of glycans. N-Glycans were then enzymatically released and subsequentially separated by porous graphitized carbon particles packed in the same device. By applying the GIG-chipLC for glycomic analysis of human sera, we identified N-glycans with 148 distinct N-glycan masses. The GIG-chipLC was used to analyze N-glycans from mouse heart tissue and serum. The extracted glycans from tissues indicated that unique un-capped N-glycans were detected in tissues that were missing from the proximal or distal serum, whereas common N-glycans from tissues and serum have mature and capped structures. The GIG-chipLC provides a simple and robust platform for glycomic analysis of complex biological and clinical samples.

Introduction:

Education and Training (in chronological order):

Undergraduate

1989       B.S.     Plant Biochemistry, Beijing University, Beijing, China

Master/graduate

1992       M.S.     Gene and Protein Engineering, Beijing University, Beijing, China

Doctoral/graduate

1999       PhD     Biochemistry, University of Pennsylvania, Philadelphia, PA

Research Scientist

2003       Postdoctoral  Protein Chemistry, Institute for Systems Biology, Seattle, WA

Professional Experience (in chronological order):

1993-1998  Research Specialist and Pre-doctoral Trainee, University of Pennsylvania, PA

1998-1999  Product Manager, New England Biolabs, Beverly, MA

1999-2001  Scientist, Cell Signaling Technology, Beverly, MA

2001-2001  Senior Scientist, Cell Signaling Technology, Beverly, MA

2001-2003  Research Scientist, Institute for Systems Biology, Seattle, WA

2003-2006  Senior Research Scientist, Institute for Systems Biology, Seattle, WA

2006-2011  Assistant Professor, Department of Pathology, Johns Hopkins University, Baltimore, MD

2011-present  Associate Professor, Department of Pathology, Johns Hopkins University, Baltimore, MD

2012-present  Director of Mass Spectrometry Core Facility, Center for Biomarker Discovery and Translation. Johns Hopkins University, Baltimore, MD

Contact:1809 Lu Wang(9620)

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