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Quantifiable fluorescent glycan microarrays

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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).

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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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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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Authors and Affiliations

  1. Department of Biochemistry, Emory University School of Medicine, O. Wayne Rollins Research Center, 1510 Clifton Road, Suite 4001, Atlanta, GA, 30322, USA

    Xuezheng Song, Baoyun Xia, Yi Lasanajak, David F. Smith & Richard D. Cummings

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  1. Xuezheng Song
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  2. Baoyun Xia
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  3. Yi Lasanajak
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  5. Richard D. Cummings
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Correspondence to Richard D. Cummings.

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Song, X., Xia, B., Lasanajak, Y. et al. Quantifiable fluorescent glycan microarrays. Glycoconj J 25, 15–25 (2008). https://doi.org/10.1007/s10719-007-9066-8

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  • DOI: https://doi.org/10.1007/s10719-007-9066-8

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