Glycobiology Advance Access originally published online on August 10, 2006
Glycobiology 2006 16(12):1171-1180; doi:10.1093/glycob/cwl038
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Engineered xyloglucan specificity in a carbohydrate-binding module
2 Department of Immunotechnology, Lund University, BMC D13, SE-221 84 Lund, Sweden;
3 School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden;
4 Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Framlington Place, Newcastle upon Tyne NE2 4HH, UK; and
5 Department of Biotechnology, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden
1 To whom correspondence should be addressed; e-mail: mats.ohlin{at}immun.lth.se
Received on November 24, 2005; revised on August 7, 2006; accepted on August 8, 2006
The field of plant cell wall biology is constantly growing and consequently so is the need for more sensitive and specific probes for individual wall components. Xyloglucan is a key polysaccharide widely distributed in the plant kingdom in both structural and storage tissues that exist in both fucosylated and non-fucosylated variants. Presently, the only xyloglucan marker available is the monoclonal antibody CCRC-M1 that is specific to terminal 
-1,2-linked fucosyl residues on xyloglucan oligo- and polysaccharides. As a viable alternative to searches for natural binding proteins or creation of new monoclonal antibodies, an approach to select xyloglucan-specific binding proteins from a combinatorial library of the carbohydrate-binding module, CBM4-2, from xylanase Xyn10A of Rhodothermus marinus is described. Using phage display technology in combination with a chemoenzymatic method to anchor xyloglucan to solid supports, the selection of xyloglucan-binding modules with no detectable residual wild-type xylan and ß-glucan-binding ability was achieved.
Key words: binding specificity / carbohydrate-binding module / molecular engineering / phage display / xyloglucan