Abstract
Plant lectin recognition of glycans was evaluated
by SPR imaging using a model array of N-biotinylated
aminoethyl glycosides of ß-D-glucose (negative control),
a-D-mannose (conA-responsive), ß-D-galactose (RCA120-
responsive) and N-acetyl-ß-D-glucosamine (WGA-responsiveresponsive) printed onto neutravidin-coated gold chips. Selective recognition of the cognate ligand was observed when RCA120 was passed over the array surface. Limited or no
binding was observed for the non-cognate ligands. SPR
imaging of an array of 40 sialylated and unsialylated
glycans established the binding preference of hSiglec7 for
a2-8-linked disialic acid structures over a2-6-sialyl-Lac-
NAcs, which in turn were recognized and bound with
greater affinity than a2-3-sialyl-LacNAcs. Affinity binding
data could be obtained with as little as 10–20 µg of lectin
per experiment. The SPR imaging technique was also able
to establish selective binding to the preferred glycan ligand
when analyzing crude culture supernatant containing 10–
20 µg of recombinant hSiglec7-Fc. Our results show that
SPR imaging provides results that are in agreement with
those obtained from fluorescence based carbohydrate arrays
but with the added advantage of label-free analysis.
Original language | English |
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Pages (from-to) | 69-74 |
Number of pages | 6 |
Journal | Glycoconjugate Journal |
Volume | 25 |
Issue number | 1 |
DOIs | |
Publication status | Published - 2008 |