Glycoconjugate Journal

, Volume 25, Issue 1, pp 15–25 | Cite as

Quantifiable fluorescent glycan microarrays

  • Xuezheng Song
  • Baoyun Xia
  • Yi Lasanajak
  • David F. Smith
  • Richard D. Cummings
Article
First Online:
Received:
Revised:
Accepted:

Abstract

A glycan microarray was developed by using 2,6-diaminopyridine (DAP) as a fluorescent linker and printing of the glycan-DAP conjugates (GDAPs) on epoxy-activated glass slides. Importantly, all coupled GDAPs showed a detectable level of concentration-dependent GDAP fluorescence under blue laser excitation (495 nm) that can be used for both grid location and on-slide quantification. A glycan array including a large number of GDAP’s derived from natural and commercially available free glycans was constructed and glycan interactions with various plant lectins were investigated. In addition, binding parameters of lectins to glycans were obtained by varying both the amount of GDAPs on the array and the lectin concentration in analyses. These data demonstrate the general utility of GDAP microarrays for functional glycomic analyses and for determining binding parameters of glycan binding proteins (GBPs).

Keywords

Carbohydrate Glycan conjugate Glycomics Microarray Fluorescence 

Abbreviations

DAP

2,6-diaminopyridine

GDAP

glycan-DAP conjugate

HPLC

high performance liquid chromatography

NHS

N-hydroxysuccinimde

RFU

relative fluorescence unit

GBP

glycan binding protein

RCA I

Ricinus communis agglutinin I

AAL

Aleuria aurantia lectin

ConA

concanavalin A

SNA

Sambucus nigra agglutinin

LTL

Lotus tetragonolobus lectin

LNnH

lacto-N-neohexaose

LNFIII

lacto-N-fucopentaose III

FITC

fluorescein isothiocyanate

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Notes

Acknowledgements

This work was supported in part by a Bridge Grant to R.D.C. from the Consortium for Functional Glycomics under NIH Grant GM62116.

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Xuezheng Song
    • 1
  • Baoyun Xia
    • 1
  • Yi Lasanajak
    • 1
  • David F. Smith
    • 1
  • Richard D. Cummings
    • 1
  1. 1.Department of BiochemistryEmory University School of MedicineAtlantaUSA

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