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

TY - JOUR

T1 - Surface plasmon resonance imaging for real-time, label-free analysis of protein interactions with carbohydrate microarrays

AU - Karamanska, Rositsa

AU - Clarke, Jonathan

AU - Blixt, Ola

AU - MacRae, James I.

AU - Zhang, Jiquan Q.

AU - Crocker, Paul R.

AU - Laurent, Nicolas

AU - Wright, Adam

AU - Flitsch, Sabine L.

AU - Russell, David A.

AU - Field, Robert A.

PY - 2008/1

Y1 - 2008/1

N2 - Plant lectin recognition of glycans was evaluated by SPR imaging using a model array of N-biotinylated aminoethyl glycosides of beta-D-glucose ( negative control), alpha-D-mannose ( conA-responsive), beta-D-galactose ( RCA(120)-responsive) and N-acetyl-beta-D-glucosamine ( WGA-responsive) printed onto neutravidin-coated gold chips. Selective recognition of the cognate ligand was observed when RCA(120) 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 alpha 2-8-linked disialic acid structures over alpha 2-6-sialyl-LacNAcs, which in turn were recognized and bound with greater affinity than alpha 2-3-sialyl-LacNAcs. Affinity binding data could be obtained with as little as 10-20 mu g 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 mu g 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.

AB - Plant lectin recognition of glycans was evaluated by SPR imaging using a model array of N-biotinylated aminoethyl glycosides of beta-D-glucose ( negative control), alpha-D-mannose ( conA-responsive), beta-D-galactose ( RCA(120)-responsive) and N-acetyl-beta-D-glucosamine ( WGA-responsive) printed onto neutravidin-coated gold chips. Selective recognition of the cognate ligand was observed when RCA(120) 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 alpha 2-8-linked disialic acid structures over alpha 2-6-sialyl-LacNAcs, which in turn were recognized and bound with greater affinity than alpha 2-3-sialyl-LacNAcs. Affinity binding data could be obtained with as little as 10-20 mu g 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 mu g 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.

KW - Carbohydrate array

KW - Surface plasmon resonance (SPR) imaging

KW - Label-free detection

KW - Plant lectin

KW - Siglec 7

KW - Immunoglobulin superfamily

KW - 2-bromoethyl glycosides

KW - Immune system

KW - Throughput

KW - Binding

KW - Oligosaccharides

KW - Recognition

KW - Lectin

KW - Inhibition

KW - Biosensors

U2 - 10.1007/s10719-007-9047-y

DO - 10.1007/s10719-007-9047-y

M3 - Article

C2 - 17574526

SN - 0282-0080

VL - 25

SP - 69

EP - 74

JO - Glycoconjugate Journal

JF - Glycoconjugate Journal

IS - 1

ER -