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A novel approach to chemical microarray using ketone-modified macromolecular scaffolds: Application in micro cell-adhesion assay

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Abstract

This paper describes a novel strategy for the preparation of chemical microarrays using macro-molecular scaffolds. The macromolecular scaffolds are first functionalized with ketone groups and compounds of interest containing an aminooxy group are conjugated onto the ketone-modified scaffolds through a chemoselective oxime ligation. The conjugate mixtures are then spotted directly onto a plastic or glass surface to form compound microarrays. Because a constant amount of scaffold is used in the presence of excess compound in the ligation reaction, the amount of compound actually immobilized per microarray spot is constant and dependent on the scaffold concentration. Using this approach, 60 different peptides were ligated to human serum albumin or agarose scaffolds, and the peptide conjugates subsequently printed on glass or polystyrene surface to form microarrays. These peptide microarrays were subsequently evaluated and optimized for binding of Jurkat leukemic cancer cells.

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References

  1. Lam, K.S. and Renil, M., From combinatorial chemistry to chemical microarray, Curr. Opin. Chem. Biol., 6 (2002) 353-8.

    PubMed  Google Scholar 

  2. Xu, Q. and Lam, K.S., Protein and chemical microarrays- Powerful tools for proteomics, J. Biomed. Biotech., 2003 (in press).

  3. Martin, B.D., Gabaer, B.P., Patterson, C.H. and Turner, D.C., Direct protein microarray fabrication using a hydrogel 'stamper', Langmuir, 14 (1998) 3971-3975.

    Google Scholar 

  4. Ge, H., UPA, a universal protein array system for quantitative detection of protein-protein, protein-DNA, protein-RNA and protein-ligand interactions, Nucleic Acids Res., 28 (2000) e3.

  5. Huang, R.-P., Detection of multiple proteins in an antibody-based protein microarray system, J. Immunol. Meth., 255 (2001) 1-13.

    Google Scholar 

  6. MacBeath, G. and Schreiber, S.L., Printing proteins as microarrays for high-throughput function determination, Science, 289 (2000) 1760-1763.

    Google Scholar 

  7. Falsey, J.R., Renil, M., Park, S., Li, S. and Lam, K.S., Peptide and small molecule microarray for high throughput cell adhesion and functional assays, Bioconjugate Chem., 12 (2001) 346-353.

    Google Scholar 

  8. Frank, R., Spot-synthesis: An easy technique for the positionally addressable, parallel chemical synthesis on a membrane support, Tetrahedron, 48 (1992) 9217.

    Google Scholar 

  9. Fodor, S.P., Read, J.L., Pirrung, M.C., Stryer, L., Lu, A.T. and Solas, D., Light-directed, spatially addressable parallel chemical synthesis, Science, 251 (1991) 767-73.

    PubMed  Google Scholar 

  10. Pease, A.C., Solas, D., Sullivan, E.J., Cronin, M.T., Holmes, C.P. and Fodor, S.P.A., Light-generated oligonucleotide arrays for rapid DNA sequence analysis, Proc. Natl. Acad. Sci. USA, 21 (1994) 5022-5026.

    Google Scholar 

  11. Singh-Gasson, S., Green, R.D., Yue, Y., Nelson, C., Blattner, F., Sussman, M.R. and Cerrina, F., Maskless fabrication of light-directed oligonucleotide microarrays using a digital micromirror array, Nat. Biotechnol., 17 (1999) 974-978.

    PubMed  Google Scholar 

  12. LeProust, E., Pellois, J.P., Yu, P., Zhang, H., Gao, X., Srivannavit, O., Gulari, E., Zhou, X., Digital light-directed synthesis. A microarray platform that permits rapid reaction optimization on a combinatorial basis, J. Comb. Chem., 2 (2000) 349-354.

    PubMed  Google Scholar 

  13. Pellois, J.P., Wang, W. and Gao, X.: Peptide synthesis based on t-Boc chemistry and solution photogenerated acids, J. Comb. Chem., 2 (2000) 355-360.

    PubMed  Google Scholar 

  14. Lin, S.C., Tseng, F.G., Huang, H.M., Huang, C.Y. and Chieng, C.C., Microsized 2D protein arrays immobilized by micro-stamps and micro-wells for disease diagnosis and drug screening, Fresenius J. Anal. Chem., 371 (2001) 202-208.

    PubMed  Google Scholar 

  15. Benters, R., Niemeyer, C.M. and Wohrle, D., Dendrimer-activated solid supports for nucleic acid and protein microarray, ChemBiochem., 2 (2001) 686-694.

    PubMed  Google Scholar 

  16. MacBeath, G., Koehler, A.N. and Schreiber, S.L., Printing small molecules as microarrays and detecting protein-ligand interactions en masse, J. Am. Chem. Soc., 121 (1999) 7967-7968.

    Google Scholar 

  17. Xu, Q. and Lam, K.S., An efficient approach to prepare glyoxylyl functionality on solid-support, Tetrahedron Lett., 43 (2002) 4435-4437.

    Google Scholar 

  18. Song, A., Wang, X., Zhang, J., Marik, J., Lebrilla, C.B. and Lam, K.S., Synthesis of hydrophilic and flexible linkers for peptide derivatization in solid phase, Bioorg. Med. Chem. Lett., 2003 (in press).

  19. Aina, O.H., Sroka, T.C., Chen, M.L., Lam, K.S., Therapeutic cancer targeting peptides, Biopolymers, 66 (2002) 184-199.

    PubMed  Google Scholar 

  20. Park, S., Renil, M., Vikstrom, B., Amro, N. and Lam, K.S., Identification of peptide ligands for a4β1 integrin receptor as potential targeting agents for non-Hodgkin's lymphoma, InM. Lebl and R.A. Houghten (Eds), Peptides: The Wave of the Future (Proceedings of the 2nd International and the 17th American Peptide Symposium), San Diego, CA, United States, 9-14 June 2001, Kluwer Academic Publishers, Dordrecht, 2002, pp. 180-182.

    Google Scholar 

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Xu, Q., Miyamoto, S. & Lam, K.S. A novel approach to chemical microarray using ketone-modified macromolecular scaffolds: Application in micro cell-adhesion assay. Mol Divers 8, 301–310 (2004). https://doi.org/10.1023/B:MODI.0000036238.42200.3d

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  • DOI: https://doi.org/10.1023/B:MODI.0000036238.42200.3d

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