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

Karamanska, R, Clarke, J, Blixt, O, MacRae, JI, Zhang, JQ, Crocker, PR, Laurent, N, Wright, A, Flitsch, SL, Russell, DA and Field, RA (2008) Surface plasmon resonance imaging for real-time, label-free analysis of protein interactions with carbohydrate microarrays. Glycoconjugate Journal, 25 (1). pp. 69-74. ISSN 0282-0080

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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.

Item Type: Article
Faculty \ School: Faculty of Science > School of Chemistry
Related URLs:
Depositing User: Rachel Smith
Date Deposited: 24 Jan 2011 10:28
Last Modified: 07 Jan 2019 14:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/19728
DOI: 10.1007/s10719-007-9047-y

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