Authors: Samnang Tep Marina Hincapie William S Hancock
Publish Date: 2012/01/26
Volume: 402, Issue: 9, Pages: 2687-2700
Abstract
The development of a general method for the purification and quantitative glycomic analysis of human plasma samples to characterize global glycosylation changes shall be presented The method involves multiple steps including the depletion of plasma via multiaffinity chromatography to remove high abundant proteins the enrichment of the lower abundant glycoproteins via multilectin affinity chromatography the isotopic derivatization of released glycans and quantitative analysis by MALDITOF MS Isotopic derivatization of glycans is accomplished using the wellestablished chemistry of reductive amination to derivatize glycans with either a light analog 12C anthranilic acid or a heavy analog 13C7 anthranilic acid which allows for the direct comparison of the alternately labeled glycans by MALDITOF MS The method displays a tenfold linear dynamic range for both neutral and sialylated glycans with subpicomolar sensitivity Additionally by using anthranilic acid a very sensitive fluorophore as the derivatization reagent the glycans can be analyzed by chromatography with fluorescence detection The utility of this methodology is highlighted by the many diseases and disorders that are known to either show or be the result of changes in glycosylation A method that provides a generic approach for sample preparation and quantitative data will help to further advance the field of glycomicsThe authors would like to thank our colleagues at Biogen Idec and the Barnett Institute of Northeastern University for their support guidance and review The authors would also like to express sincere appreciation to the NIH Grant No U01CA128427
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