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Fabrication of Carbohydrate Microarrays by Boronate Formation

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Small Molecule Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1518))

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Abstract

The interactions between soluble carbohydrates and/or surface displayed glycans and protein receptors are essential to many biological processes and cellular recognition events. Carbohydrate microarrays provide opportunities for high-throughput quantitative analysis of carbohydrate–protein interactions. Over the past decade, various techniques have been implemented for immobilizing glycans on solid surfaces in a microarray format. Herein, we describe a detailed protocol for fabricating carbohydrate microarrays that capitalizes on the intrinsic reactivity of boronic acid toward carbohydrates to form stable boronate diesters. A large variety of unprotected carbohydrates ranging in structure from simple disaccharides and trisaccharides to considerably more complex human milk and blood group (oligo)saccharides have been covalently immobilized in a single step on glass slides, which were derivatized with high-affinity boronic acid ligands. The immobilized ligands in these microarrays maintain the receptor-binding activities including those of lectins and antibodies according to the structures of their pendant carbohydrates for rapid analysis of a number of carbohydrate-recognition events within 30 h. This method facilitates the direct construction of otherwise difficult to obtain carbohydrate microarrays from underivatized glycans.

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References

  1. Dwek RA (1996) Glycobiology: toward understanding the function of sugars. Chem Rev 96:683–720

    Article  CAS  PubMed  Google Scholar 

  2. Helenius A, Abei M (2001) Intracellular functions of N-linked glycans. Science 291:2364–2369

    Article  CAS  PubMed  Google Scholar 

  3. Marth JD, Grewal PK (2008) Mammalian glycosylation in immunity. Nat Rev Immunol 8:874–887

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. van Kooyk Y, Rabinovich GA (2008) Protein-glycan interactions in the control of innate and adaptive immune responses. Nat Immunol 9:593–601

    Article  PubMed  Google Scholar 

  5. Lee YC, Lee RT (1995) Carbohydrate-protein interactions: basis for glycobiology. Acc Chem Res 28:321–327

    Article  CAS  Google Scholar 

  6. Imberty A, Varrot A (2008) Microbial recognition of human cell surface glycoconjugates. Curr Opin Struct Biol 18:567–576

    Article  CAS  PubMed  Google Scholar 

  7. Scanlan CN, Offer J, Zitzmann N, Dwek RA (2007) Exploiting the defensive sugars of HIV-1 for drug and vaccine design. Nature 446:1038–1045

    Article  CAS  PubMed  Google Scholar 

  8. Kiessling LL, Splain RA (2010) Chemical approaches to glycobiology. Annu Rev Biochem 79:619–653

    Article  CAS  PubMed  Google Scholar 

  9. Rillahan CD, Paulson JC (2011) Glycan microarrays for decoding the glycome. Annu Rev Biochem 80:797–823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. McCoy JP, Varani J, Goldstein IJ (1983) Enzyme-linked lectin assay (ELLA): use of alkaline phosphatase-conjugated Griffonia simplicifolia B4 isolectin for the detection of alpha-D-galactopyranosyl end groups. Anal Biochem 130:437–444

    Article  CAS  PubMed  Google Scholar 

  11. Lis H, Sharon N (1972) Lectins: cell-agglutinating and sugar-specific proteins. Science 177:949–959

    Article  PubMed  Google Scholar 

  12. Dam TK, Brewer CF (2002) Thermodynamic studies of lectin-carbohydrate interactions by isothermal titration calorimetry. Chem Rev 102:387–430

    Article  CAS  PubMed  Google Scholar 

  13. Duverger E, Frison N, Roche AC, Monsingny M (2003) Carbohydrate-lectin interactions assessed by surface plasmon resonance. Biocheimie 85:167–179

    Article  CAS  Google Scholar 

  14. Kitova EN, Kitov PI, Bundle DR, Klassen JS (2001) The observation of multivalent complexes of Shiga-like toxin with globotrioside and the determination of their stoichiometry by nanoelectrosray Fourier-transform ion cyclotron resonance mass spectroscopy. Glycobiology 11:605–611

    Article  CAS  PubMed  Google Scholar 

  15. Adak AK, Li BY, Lin CC (2015) Advances in multifunctional glycosylated nanomaterials: preparation and applications in glycoscience. Carbohydr Res 405:2–12

    Article  CAS  PubMed  Google Scholar 

  16. Adak AK, Lin HJ, Lin CC (2014) Multivalent glycosylated nanoparticles for studying carbohydrate-protein interactions. Org Biomol Chem 12:5563–5573

    Article  CAS  PubMed  Google Scholar 

  17. Fazio F, Bryan MC, Blixt O, Paulson JC, Wong CH (2002) Synthesis of sugar arrays in microtiter plate. J Am Chem Soc 124:14397–14402

    Article  CAS  PubMed  Google Scholar 

  18. Park S, Shin I (2002) Fabrication of carbohydrate chips for studying protein-carbohydrate interactions. Angew Chem Int Ed 41:3180–3182

    Article  CAS  Google Scholar 

  19. Houseman BT, Mrksich M (2002) Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. Chem Biol 9:443–454

    Article  CAS  PubMed  Google Scholar 

  20. Fukui S, Feizi T, Galustian C, Lawson AM, Chai W (2002) Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Nat Biotechnol 20:1011–1017

    Article  CAS  PubMed  Google Scholar 

  21. Wang D, Liu S, Trummer BJ, Deng C, Wang A (2002) Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells. Nat Biotechnol 20:275–281

    Article  CAS  PubMed  Google Scholar 

  22. Liang PH, Wu CY, Greenberg WA, Wong CH (2008) Glycan arrays: biological and medical applications. Curr Opin Chem Biol 12:86–92

    Article  CAS  PubMed  Google Scholar 

  23. Park S, Gildersleeve JC, Blixt O, Shin I (2013) Carbohydrate microarrays. Chem Soc Rev 42:4310–4326

    Article  CAS  PubMed  Google Scholar 

  24. Hsiao HY, Chen ML, Wu HT, Huang LD, Chien WT, Yu CC, Jan FD, Sahabuddin S, Chang TC, Lin CC (2008) Fabrication of carbohydrate microarrays through boronate formation. Chem Commun 47:1187–1189

    Article  Google Scholar 

  25. James TD, Sandanayake KRAS, Shinkai S (1996) Saccharide sensing with molecular receptors based on boronic acid. Angew Chem Int Ed 35:1911–1922

    Article  CAS  Google Scholar 

  26. Yang YL, Lee YP, Yang YL, Lin PC (2014) Traceless labeling of glycoproteins and its application to the study of glycoprotein-protein interactions. ACS Chem Biol 9:390–397

    Article  CAS  PubMed  Google Scholar 

  27. Chen ML, Adak AK, Yeh NC, Yang WB, Chuang YJ, Wong CH, Hwang KC, Reuben JR, Hsieh SL, Lin CC (2008) Fabrication of an oriented Fc-fused lectin microarray through boronate formation. Angew Chem Int Ed 47:8627–8630

    Article  CAS  Google Scholar 

  28. Adak AK, Li BY, Huang LD, Lin TW, Chang CT, Hwang KC, Lin CC (2014) Fabrication of antibody microarrays by light-induced covalent and oriented immobilization. ACS Appl Mater Interfaces 6:10452–10460

    Article  CAS  PubMed  Google Scholar 

  29. Huang LD, Adak AK, Yu CC, Hsiao WC, Lin HJ, Chen ML, Lin CC (2015) Fabrication of highly stable glyco-gold nanoparticles and development of a glyco-gold nanoparticle-based oriented immobilized antibody microarray for lectin (GOAL) assay. Chem Eur J 21:3956–3967

    Article  CAS  PubMed  Google Scholar 

  30. Lin PC, Chen SH, Wang KY, Chen ML, Adak AK, Hwu RJ, Chen YJ, Lin CC (2009) Fabrication of oriented antibody-conjugated magnetic nanoprobes and their immunoaffinity application. Anal Chem 81:8774–8782

    Article  CAS  PubMed  Google Scholar 

  31. Lu YW, Chien CW, Lin PC, Huang LD, Chen CY, Han CL, Khoo KH, Lin CC, Chen YJ (2013) BAD-lectins: boronic acid-decorated lectins with enhanced binding affinity for the selective enrichment of glycoproteins. Anal Chem 85:8268–8276

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We are pleased to acknowledge the financial support of National Tsing Hua University, Academia Sinica, and Ministry of Science and Technology, Taiwan.

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Authors and Affiliations

  1. Department of Chemistry, National Tsing Hua University, 101, Sec 2 Kuang-Fu Rd, Hsinchu, 30013, Taiwan

    Avijit K. Adak, Ting-Wei Lin, Ben-Yuan Li & Chun-Cheng Lin

Authors
  1. Avijit K. Adak
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  2. Ting-Wei Lin
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  3. Ben-Yuan Li
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  4. Chun-Cheng Lin
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Corresponding authors

Correspondence to Avijit K. Adak or Chun-Cheng Lin .

Editor information

Editors and Affiliations

  1. DSO National Laboratories, Defence Medical and Environmental Resear, Singapore, Singapore

    Mahesh Uttamchandani

  2. Department of Chemistry Faculty of Science, National University of Singapore, Singapore, Singapore

    Shao Q. Yao

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Adak, A.K., Lin, TW., Li, BY., Lin, CC. (2017). Fabrication of Carbohydrate Microarrays by Boronate Formation. In: Uttamchandani, M., Yao, S. (eds) Small Molecule Microarrays. Methods in Molecular Biology, vol 1518. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6584-7_4

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  • DOI: https://doi.org/10.1007/978-1-4939-6584-7_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6582-3

  • Online ISBN: 978-1-4939-6584-7

  • eBook Packages: Springer Protocols

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