Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research

Abstract

Background: Microarrays of plant-derived oligosaccharides are potentially powerful tools for the high throughput discovery and screening of antibodies, enzymes, and carbohydrate-binding proteins. Results: Oligosaccharide microarrays were produced, and their utility was demonstrated in several applications. Conclusion:A new generation of oligosaccharide microarrays will make an important contribution to plant glycomic research. Significance: High throughput screening technology enables the more effective production of carbohydrate active enzymes and molecular probes.

Escherichia coli Common Pilus (ECP) Targets Arabinosyl Residues in Plant Cell Walls to Mediate Adhesion to Fresh Produce Plants

“…To confirm the specificity, two arabinose-specific antibodies were used, LM6 and LM13. LM6 recognizes linear chains of (135)-␣-arabinan from pectin and some AGPs (39,43), whereas LM13 is specific to linear chains of (135)-␣-arabinan from pectin but importantly only to oligosaccharides longer than arabinotriose (38,44,45). As expected, LM6 recognized arabinose residues on the glycan arrays of all sizes, whereas LM13 recognized residues from arabinotriose to arabinoheptaose present on the array (Fig.…”

“…Glycan Array Printing and Screening-Glycan array printing was performed as described (38), and arrays were screened as described previously (4). For detailed information on all oligosaccharide samples, see Table 4.…”

Understanding plant cell-wall remodelling during the symbiotic interaction between Tuber melanosporum and Corylus avellana using a carbohydrate microarray

“…Cell wall polymers were extracted sequentially from 10 mg of AIR with 300 μl of 50 mM diamino-cyclo-hexane-tetra-acetic acid (CDTA), pH 7.5 and 4 M NaOH with 0.1 % v/v NaBH4 and spotted on a nitrocellulose membrane with a pore size of 0.45 μm (Whatman, Maidstone, UK) using an Arrayjet Sprint (Arrayjey, Roslin, UK). Each sample was printed with two technical replicates and four dilutions, and probed as described in Pedersen et al (2012). The arrays were scanned using a flatbed scanner (CanoScan 8800 F, Canon, Søborg, Denmark) at 2400 dpi and quantified using Array-Pro Analyzer 6.3 (Media Cybernetics, Rockville, MD, USA).…”

Monitoring Polysaccharide Dynamics in the Plant Cell Wall

“…The de novo synthesis of various oligosaccharides has enabled the characterization of xylan-directed mAbs (Schmidt et al, 2015). While microarrays with plant polysaccharides (Moller et al, 2008) or oligosaccharides (Pedersen et al, 2012) have been probed previously with mAbs (Pattathil et al, 2010), arrays of synthetic glycans have an exceptional purity and facilitate the precise mapping of the epitopes recognized by molecular probes. A comprehensive screen of 209 walldirected mAbs against 88 synthetic glycans, including 22 xylan oligosaccharides, identified specific epitopes for 78 probes that were uncharacterized previously (Ruprecht et al, 2017).…”