Use of the Avidin-Biotin Complex for Specific Immobilization of Xyloglucan Polysaccharides1

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Abstract
Several xyloglucan polysaccharides were biotinylated through their galactosyl residues, by combination of partial galactose oxidation and biotinamidocaproylhydrazide coupling. They were shown to easily bind to avidin activated ELISA plates by specific recognition of plant lectins Ulex europeaus (UEA-I) and Ricinus communis agglutinin (RCA-I). Subsequent chemical reduction of aldehyde groups after biotinylation was shown to improve immobilisation. UEA-I bound to cell wall xyloglucan but not to tamarind seed xyloglucan, due to the lack of fucose in the latter. RCA-I bound to all xyloglucans we tested, however, a higher affinity was observed with seed xyloglucan, suggesting it specifically clusters with the polysaccharide. The usefulness of using biotinylated xyloglucans for antibodies characterization was investigated.