Glycome profiling using glycan-directed monoclonal antibodies: Applications to plant cell wall/ biomass characterization

Funding

Sivakumar Pattathil1, Michael G. Hahn2, Heather McCormick1, Jeffrey Miller2, Virginia Brown2, Ajaya Biswal2, Trina Saffold2, Malcolm O'Neill2, Debra Mohnen2, William S. York2, Jaclyn D. DeMartini3, Charles Wyman4, Michael W. W. Adams5, Irina Kataeva6, Richard A. Dixon7, Huanzhong Wang7 and Fang Chen7, (1)Complex Carbohydrate Research Center, University of Georgia, Athens, GA, (2)University of Georgia, Athens, GA, (3)Chemical and Environmental Engineering, University of California Riverside, Riverside, CA, (4) University of California Riverside, Riverside, CA, (5)Biochemistry and Molecular Biology, University of Georgia, Athens, GA, (6)Department of Biochemistry, University of Georgia, Athens, GA, (7)Samuel Roberts Noble Foundation, Ardmore, OK Plant biomass is the major source for lingo-cellulosic bioenergy. Plant cell walls form the major part of the plant biomass. Research on plant cell wall/biomass is challenging owing to the complexity of the cell walls. Thus, one of the focus areas of bio-energy research has been the development of advanced tools that enable easy, rapid, high-throughput and reliable characterization of plant cell walls/ biomass. Cell wall-directed monoclonal antibodies have now emerged as powerful probes to assist in the detailed structural analysis of plant cell walls. The worldwide collection of plant cell wall glycan-directed monoclonal antibodies is now sufficiently large and comprehensive to be used to monitor the majority of cell wall glycans. Here we report an ELISA-based method, Glycome Profiling, that employs a toolkit of 155 monoclonal antibodies (recognizing most major classes of plant polysaccharides) for characterizing diverse plant cell walls. The method involves sequential extraction of cell wall materials with increasingly harsh reagents to obtain fractions enriched in diverse wall polysaccharides, such as pectins, arabinogalactans, xylans, xyloglucans, and mannans. Each of these fractions is subjected to ELISA against the toolkit of antibodies and the binding responses are depicted as heat maps. The studies presented here outline multiple applications of Glycome Profiling to plant cell wall/biomass characterization and utilization. [This work is supported by a grant from US Department of Energy BioEnergy Science Center and grant DE-PS02-06ER64304. The BioEnergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Generation of the monoclonal antibodies used in this work was supported by the NSF Plant Genome Program (DBI-0421683)]