《631_ftp》.pdf

Proteomics 2011, 11, 631–643 DOI 10.1002/pmic.201000517 631 REVIEW Glycan analysis by modern instrumental methods Martin Pabst and Friedrich Altmann Department of Chemistry, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria The oligosaccharides attached to proteins or lipids are among the most challenging analytical Received: August 17, 2010 tasks due to their complexity and variety. Knowing the genes and enzymes responsible for Revised: November 2, 2010 their biosynthesis, a large but not unlimited number of different structures and isomers of Accepted: November 3, 2010 such glycans can be imagined. Understanding of the biological role of structural variations requires the ability to unambiguously determine the identity and quantity of all glycan species. Here, we examine, which analytical strategies – with a certain high-throughput potential – may come near this ideal. After an expose of the relevant techniques, we try to depict how analytical raw data are translated into structural assignments using retention times, mass and fragment spectra. A method’s ability to discriminate between the many conceivable isomeric structures together with the time, effort and sample amount needed for that purpose is suggested as a criterion for the comparative assessment of approaches and their evolutionary stages. Keywords: Glycomics / Glycoprotein / Glycoproteomics / Mass spectrometry / Oligosaccharide / Porous graphitic carbon 1 Introduction procal relation between complexity of the analyte and rele- vance of the biological role. DNA for example manages its Some 50 years