Glycoconjugate Journal

, Volume 25, Issue 1, pp 75–84 | Cite as

Serum antibody screening by surface plasmon resonance using a natural glycan microarray

  • Arjen R. de BoerEmail author
  • Cornelis H. Hokke
  • André M. Deelder
  • M. Wuhrer


A surface plasmon resonance (SPR) based natural glycan microarray was developed for screening of interactions between glycans and carbohydrate-binding proteins (CBPs). The microarray contained 144 glycan samples and allowed the real-time and simultaneous screening for recognition by CBPs without the need of fluorescent labeling. Glycans were released from their natural source and coupled by reductive amination with the fluorescent labels 2-aminobenzamide (2AB) or anthranilic acid (AA) followed by high-performance liquid chromatography (HPLC) fractionation making use of the fluorescent tag. The released and labeled glycans, in addition to fluorescently labeled synthetic glycans and (neo)glycoproteins, were printed on an epoxide-activated chip at fmol amounts. This resulted in covalent immobilization, with the epoxide groups forming covalent bonds to the secondary amine groups present on the fluorescent glycoconjugates. The generated SPR glycan array presented a subset of the glycan repertoire of the human parasite Schistosoma mansoni. In order to demonstrate the usefulness of the array in the simultaneous detection of glycan-specific serum antibodies, the anti-glycan antibody profiles from sera of S. mansoni-infected individuals as well as from non-endemic uninfected controls were recorded. The SPR screening was sensitive for differences between infection sera and control sera, and revealed antibody titers and antibody classes (IgG or IgM). All SPR analyses were performed with a single SPR array chip, which required regeneration and blocking of the chip before the application of a serum sample. Our results indicate that SPR-based arrays constructed from glycans of natural or synthetic origin, pure or as mixture, can be used for determining serum antibody profiles as possible markers for the infection status of an individual.


Carbohydrate-binding protein detection Glycan microarray Natural glycans Serum screening Surface plasmon resonance 





anthranilic acid


circulating anodic antigen, -6)[GlcA(β1–3)]GalNAc(β1-


carbohydrate-binding protein












hydrophilic interaction liquid chromatography


high-performance liquid chromatography


keyhole limpet hemocyanin






Lewis X, Gal(β1–4)[Fuc(α1–3)]GlcNAc


matrix-assisted laser desorption/ionization-time of flight-mass spectrometry


partial least square for discriminant analysis


reversed phase


surface plasmon resonance


Schistosoma mansoni



We would like to thank Carolien A.M. Koeleman for excellent technical assistance, and we appreciate the support of Dr. Marco R. Bladergroen, Dr. Alex A. Henneman, Dr. Alexandra van Remoortere, L. Renee Ruhaak and Dr. Govert J. van Dam. A. Sylvan, E. Verschuuren and P. Lidén (GE Healthcare) are gratefully acknowledged for help and cooperation. This study was supported by the Netherlands Genomics Initiative (Horizon Breakthrough Project 050-71-302) and GE Healthcare.


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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Arjen R. de Boer
    • 1
    Email author
  • Cornelis H. Hokke
    • 1
  • André M. Deelder
    • 1
  • M. Wuhrer
    • 1
  1. 1.Biomolecular Mass Spectrometry Unit, Department of Parasitology, Center of Infectious DiseasesLeiden University Medical CenterRC LeidenThe Netherlands

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