Surface plasmon resonance imaging for real-time, label-free analysis of protein interactions with carbohydrate microarrays

Rositsa Karamanska, Jonathan Clarke, Ola Blixt, James I. MacRae, Jiquan Q. Zhang, Paul R. Crocker, Nicolas Laurent, Adam Wright, Sabine L. Flitsch, David A. Russell, Robert A. Field

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88 Citations (Scopus)

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 languageEnglish
Pages (from-to)69-74
Number of pages6
JournalGlycoconjugate Journal
Volume25
Issue number1
DOIs
Publication statusPublished - 2008

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