Abstract
A method for the preparation of carbohydrate microarrays inside 96-well polystyrene microtiter plates is described. The key step in this strategy represents the synthesis of carbohydrate–dextran conjugates by copper (I)-catalyzed [3 + 2] cycloaddition between alkyne carbohydrate derivative and a specially designed azido dextran polymer. The conjugates adsorb efficiently on polystyrene surface and can be printed inside 96-well plates using a non-contact piezoelectric microarrayer. Model interactions with a selection of lectins (concanavalin A, wheat germ agglutinin, Erythrina Cristagalli) display the efficiency of the immobilization method, its reproducibility and the specificity of biomolecular interactions occurring at the polystyrene–water interface.
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References
Love, K. R., and Seeberger, P. H. (2002) Carbohydrate arrays as tools for glycomics. Angew Chem Int Ed Engl 41, 3583–6, 3513.
Leinonen, M., and Frasch, C. E. (1982) Class-specific antibody response to group B Neisseria meningitidis capsular polysaccharide: use of polylysine precoating in an enzyme-linked immunosorbent assay. Infect Immun 38, 1203–7.
Marson, A., Robinson, D. E., Brookes, P. N., Mulloy, B., Wiles, M., Clark, S. J., Fielder, H. L., Collinson, L. J., Cain, S. A., Kielty, C. M., McArthur, S., Buttle, D. J., Short, R. D., Whittle, J. D., and Day, A. J. (2009) Development of a microtiter plate-based glycosaminoglycan array for the investigation of glycosaminoglycan-protein interactions. Glycobiology 19, 1537–46.
Zielen, S., Broker, M., Strnad, N., Schwenen, L., Schon, P., Gottwald, G., and Hofmann, D. (1996) Simple determination of polysaccharide specific antibodies by means of chemically modified ELISA plates. J Immunol Methods 193, 1–7.
Gehring, A. G., Albin, D. M., Reed, S. A., Tu, S. I., and Brewster, J. D. (2008) An antibody microarray, in multiwell plate format, for multiplex screening of foodborne pathogenic bacteria and biomolecules. Anal Bioanal Chem 391, 497–506.
Cherniak, R., Cheeseman, M. M., Reyes, G. H., Reiss, E., and Todaro, F. (1988) Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay. Diagn Clin Immunol 5, 344–8.
Gray, B. M. (1979) ELISA methodology for polysaccharide antigens: protein coupling of polysaccharides for adsorption to plastic tubes. J Immunol Methods 28, 187–92.
Fazio, F., Bryan, M. C., Blixt, O., Paulson, J. C., and Wong, C. H. (2002) Synthesis of sugar arrays in microtiter plate. J Am Chem Soc 124, 14397–402.
Satoh, A., Fukui, E., Yoshino, S., Shinoda, M., Kojima, K., and Matsumoto, I. (1999) Comparison of methods of immobilization to enzyme-linked immunosorbent assay plates for the detection of sugar chains. Anal Biochem 275, 231–5.
Dyukova, V. I., Shilova, N. V., Galanina, O. E., Rubina, A. Y., and Bovin, N. V. (2006) Design of carbohydrate multiarrays. Biochim Biophys Acta 1760, 603–9.
Graves, H. C. (1988) The effect of surface charge on non-specific binding of rabbit immunoglobulin G in solid-phase immunoassays. J Immunol Methods 111, 157–66.
Lee, M. R., and Shin, I. (2005) Facile preparation of carbohydrate microarrays by site-specific, covalent immobilization of unmodified carbohydrates on hydrazide-coated glass slides. Org Lett 7, 4269–72.
Shin, I., Park, S., and Lee, M. R. (2005) Carbohydrate microarrays: an advanced technology for functional studies of glycans. Chemistry 11, 2894–901.
Park, S., and Shin, I. (2002) Fabrication of carbohydrate chips for studying protein-carbohydrate interactions. Angew Chem Int Ed Engl 41, 3180–2.
Fukui, S., Feizi, T., Galustian, C., Lawson, A. M., and Chai, W. (2002) Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Nat Biotechnol 20, 1011–7.
Wang, D., Liu, S., Trummer, B. J., Deng, C., and Wang, A. (2002) Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells. Nat. Biotechnol. 20, 275–81.
Carion, O., Lefebvre, J., Dubreucq, G., Dahri-Correia, L., Correia, J., and Melnyk, O. (2006) Polysaccharide microarrays for polysaccharide-platelet-derived-growth-factor interaction studies. Chembiochem 7, 817–26.
Huynh, R., Chaubet, F., and Jozefonvicz, J. (2001) Anticoagulant properties of dextranmethylcarboxylate benzylamide sulfate (DMCBSu); a new generation of bioactive functionalized dextran. Carbohydr Res 332, 75–83.
Rostovtsev, V. V., Green, L. G., Fokin, V. V., and Sharpless, K. B. (2002) A stepwise huisgen cycloaddition process: copper(I)-catalyzed regioselective “ligation” of azides and terminal alkynes. Angew Chem Int Ed Engl 41, 2596–9.
Tornoe, C. W., Christensen, C., and Meldal, M. (2002) Peptidotriazoles on Solid Phase: [1,2,30-Triazoles by Regiospecific Copper(I)-Catalyzed 1,3-Dipolar Cycloadditions of Terminal Alkynes to Azides.. J. Org. Chem. 67, 3057–3064.
Carboni, B., Benalil, A., and Vaultier, M. (1993) Aliphatic amino azides as key building blocks for efficient polyamine syntheses. J. Org. Chem. 58, 736–3741.
Tamanini, E., Rigby, S. E., Motevalli, M., Todd, M. H., and Watkinson, M. (2009) Responsive metal complexes: a click-based “allosteric scorpionate” complex permits the detection of a biological recognition event by EPR/ENDOR spectroscopy. Chem. Eur. J. 15, 3720–8.
Wan, Q., Chen, J., Chen, G., and Danishefsky, S. L. (2006) A potentially valuable advance in the synthesis of carbohydrate-based anticancer vaccines through extended cycloaddition chemistry. J. Org. Chem. 71, 8244–8249.
Roy, B., and Mukhopadhyay, B. (2007) Sulfuric acid immobilized on silica: an excellent catalyst for Fischer type glycosylation. Tetrahedron Lett. 48, 3783–3787.
Acknowledgments
We gratefully acknowledge financial support from CNRS, Université de Lille Nord de France, Institut Pasteur de Lille, IFR 142, Région Nord Pas de Calais, the European Community (FEDER), Endotis Pharma, Inc., and from Cancéropôle Nord-Ouest. This research was performed using the Chemistry Systems Biology platform (http://csb.ibl.fr).
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Ebran, JP., Dendane, N., Melnyk, O. (2012). Carbohydrate Microarrays in 96-Well Polystyrene Microtiter Plates. In: Chevolot, Y. (eds) Carbohydrate Microarrays. Methods in Molecular Biology, vol 808. Humana Press. https://doi.org/10.1007/978-1-61779-373-8_25
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DOI: https://doi.org/10.1007/978-1-61779-373-8_25
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