tailieunhanh - Báo cáo khoa học: Glycomics-based analysis of chicken red blood cells provides insight into the selectivity of the viral agglutination assay

Agglutination of red blood cells (RBCs), including chicken RBCs (cRBCs), has been used extensively to estimate viral titer, to screen glycan-receptor binding preference, and to assess the protective response of vaccines. Although this assay enjoys widespread use, some virus strains do not agglutinate RBCs. | IFEBS Journal Glycomics-based analysis of chicken red blood cells provides insight into the selectivity of the viral agglutination assay Udayanath Aich1 Nia Beckley1 Zachary Shriver1 Rahul Raman1 Karthik Viswanathan1 Sven Hobbie2 and Ram Sasisekharan1 2 1 Harvard-MIT Division of Health Sciences Technology the Koch Institute for Integrative Cancer Research and the Department of Biological Engineering Massachusetts Institute of Technology Cambridge MA USA 2 Singapore-MIT Alliance for Research and Technology Centre for Life Sciences Singapore Keywords glycans influenza mass spectrometry nuclear magnetic resonance red blood cells Correspondence R. Sasisekharan 77 Massachusetts Avenue E25-519 Cambridge MA 02139 USA Fax 1 617 258 9409 Tel 1 617 258 9494 E-mail rams@ Received 2 December 2010 revised 3 February 2011 accepted 11 March 2011 doi Agglutination of red blood cells RBCs including chicken RBCs cRBCs has been used extensively to estimate viral titer to screen glycan-receptor binding preference and to assess the protective response of vaccines. Although this assay enjoys widespread use some virus strains do not agglutinate RBCs. To address these underlying issues and to increase the usefulness of cRBCs as tools for studying viruses such as influenza we analyzed the cell surface N-glycans of cRBCs. On the basis of the results obtained from complementary analytical strategies including MS 1D and 2D-NMR spectroscopy exoglycosidase digestions and HPLC profiling we report the major glycan structures present on cRBCs. By comparing the glycan structures of cBRCs with those of representative human upper respiratory cells we offer a possible explanation for the fact that certain influenza strains do not agglutinate cRBCs using specific human-adapted influenza hemagglutinins as examples. Finally recent understanding of the role of various glycan structures in high affinity binding to influenza hemagglutinins provides context to our .