FEATURE RESEARCH
Development of MS-based analytical methods for glycomics & glycoproteomics
BACKGROUND OF RESEARCH PROF. HYUN JOO AN Dr Hyun Joo An is the director of the Asia Glycomics Reference Site (AGRS), which develops and validates new analytical platforms for glycomic and glycoproteomic Department: AGRS, GRAST, analysis in collaboration with government agencies and regional industry. Since Chungnam National University 2011, Dr An has been an associate professor at Chungnam National University. She Phone Number: 82-42-821-8547 received her BS and MS degrees from Inha University (Incheon, Korea) and her PhD Email: [email protected] from the University of California at Davis (United States). Dr. An’s research focuses on bioanalytical mass spectrometry, with applications to glycomics, proteomics, and glycoproteomics. Her specific research interests include bioanalytical method development, biopharmaceutical characterization, disease biomarker discovery, clinical glycomics, and glycobiology. She has authored and co-authored over a 60 peer-reviewed publications on these subjects, and holds multiple related patents. The Asia Glycomics Reference Site (AGRS), under the directorship of Prof. Hyun Joo An, specializes in mass spectrometric characterization of glycans, glycoproteins, and proteins. The AGRS develops and validates new analytical platforms for glycomic and glycoproteomic analysis, using cutting-edge facilities and equipment. In addition to providing core glyco-analytical support to biopharmaceutical companies as well as governmental regulatory agencies, the AGRS conducts basic glycobiological research in collaboration with academic labs around the world.
OBJECTIVES OF RESEARCH - Efficient and selective method to capture O-glycans from complex mixture - Isomer-specific glycan separation and quantitation - Selective enrichment of phosphoglycans using newly designed LC/MS chip RESULT OF RESEARCH that can be applied in glycoproteomics and glycomic research. These new strategies will help us gain a greater understanding Despite recent advances, site-specific profiling of protein of an important yet largely unexplored portion of our biology: the glycosylation remains a significant analytical challenge for glycome. conventional proteomic methodology. To alleviate the issue, we propose glyco-analytical multispecific proteolysis (Glyco- AMP) as a strategy for glycoproteomic characterization. REFERENCE OF PUBLICATION Glyco-AMP consists of rapid, in-solution digestion of 1. Hua, S., Hu, C.Y., Kim, B.J., Totten, S.M., Oh, M.J., Yun, an analyte glycoprotein (or glycoprotein mixture) by a N.Y., Nwosu, C.C., Yoo, J.S., Lebrilla, C.B., An, H.J., multispecific protease (or protease cocktail). Resulting “Glyco-Analytical Multispecific Proteolysis (Glyco-AMP): glycopeptides are chromatographically separated by isomer- A Simple Method for Detailed and Quantitative specific porous graphitized carbon nano-LC, quantified by Glycoproteomic Characterization”, J. Proteome Res, 2013. high-resolution MS, and structurally elucidated by MS/MS. To demonstrate the consistency and customizability of 2. Strum, J.S., Nwosu, C.C., Hua, S., Kronewitter, S.R., Glyco-AMP methodology, the glyco-analytical performances Seipert, R.R., Bachelor, R.J., An, H.J., Lebrilla, C.B., of multispecific proteases subtilisin, pronase, and proteinase “Automated Assignments of N- and O-Site Specific K were characterized in terms of quantitative accuracy, Glycosylation with Extensive Glycan Heterogeneity of sensitivity, and digestion kinetics. Glyco-AMP was shown be Glycoprotein Mixtures”, Anal. Chem, 2013, 85, 5666−5675. effective on glycoprotein mixtures as well as glycoproteins 3. Hua, S., Oh, M.J., An, H.J., “Multi-Level Characterization with multiple glycosylation sites, providing detailed, of Protein Glycosylation”, Mass Spectrom. Lett, 2013, quantitative, site- and structure-specific information about 4(1), 10–17. protein glycosylation. 4. Nwosu, C.C., Seipert, R.R., Strum, J.S., Hua, S., An, H.J., The AGRS has developed new and improved analytical Zivkovic, A.M., German, B.J., Lebrilla, C.B., “Simultaneous techniques for characterization of the glycoproteome, and Extensive Site-specific N- and O-Glycosylation including (in order of increasing complexity and information Analysis in Protein Mixtures”, J. Proteome Res, 2011, 10, content) preliminary site mapping, compositional glycan 2612–2624. profiling, isomer-specific glycan profiling, glycosite-specific glycopeptide profiling, and glycoproteomic profiling using 5. Hua, S., Lebrilla, C.B., An, H.J., “Application of Agilent 6500 Series Accurate-Mass Quadrupole Time-of-Flight nano-LC-based glycomics towards biomarker discovery”, (Q-TOF) LC/MS interfaced with 1260 HPLC Chip cube system Bioanalysis, 2011, 3(22), 2573–2585.