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

Plant lectin recognition of glycans was evaluated by SPR imaging using a model array of N-biotinylated aminoethyl glycosides of β-d-glucose (negative control), α-d-mannose (conA-responsive), β-d-galactose (RCA₁₂₀-responsive) and N-acetyl-β-d-glucosamine (WGA-responsive) printed onto neutravidin-coat...

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Bibliographic Details
Published in:Glycoconjugate journal 2008, Vol.25 (1), p.69-74
Main Authors: Karamanska, Rositsa, Clarke, Jonathan, Blixt, Ola, MacRae, James I, Zhang, Jiquan Q, Crocker, Paul R, Laurent, Nicolas, Wright, Adam, Flitsch, Sabine L, Russell, David A, Field, Robert A
Format: Article
Language:English
Subjects:
Biochemistry
Biomedical and Life Sciences
Biotinylation
Carbohydrate array
Carbohydrate Metabolism
Carbohydrates
Carbohydrates - analysis
Glycomics - methods
Label-free detection
Life Sciences
Microarray Analysis - methods
Pathology
Plant lectin
Protein Binding
Proteins
Proteins - analysis
Proteins - metabolism
Siglec 7
Studies
Surface plasmon resonance (SPR) imaging
Surface Plasmon Resonance - methods
Time Factors
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Summary:Plant lectin recognition of glycans was evaluated by SPR imaging using a model array of N-biotinylated aminoethyl glycosides of β-d-glucose (negative control), α-d-mannose (conA-responsive), β-d-galactose (RCA₁₂₀-responsive) and N-acetyl-β-d-glucosamine (WGA-responsive) 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 α2-8-linked disialic acid structures over α2-6-sialyl-LacNAcs, which in turn were recognized and bound with greater affinity than α2-3-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.
ISSN:0282-0080
1573-4986
DOI:10.1007/s10719-007-9047-y