Abstract
Peptide microarrays are useful tools for characterizing the humoral response against methylated antigens. They are usually prepared by printing unmodified and methylated peptides on substrates such as functionalized microscope glass slides. The preferential capture of antibodies by methylated peptides suggests the specific recognition of methylated epitopes. However, unmodified peptide epitopes can be masked due to their interaction with the substrate. The accessibility of unmodified peptides and thus the specificity of the recognition of methylated peptide epitopes can be probed using the in situ methylation procedure described here. Alternately, the in situ methylation of peptide microarrays allows probing the presence of antibodies directed toward methylated epitopes starting from easy-to-make and cost-effective unmodified peptide libraries. In situ methylation was performed using formaldehyde in the presence of sodium cyanoborohydride and nickel chloride. This chemical procedure converts lysine residues into mono- or dimethyl lysines.
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Acknowledgments
We gratefully acknowledge the 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), and from Cancéropôle Nord-Ouest. This research was performed using the Chemistry Systems Biology platform (http://csb.ibl.fr).
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Desmet, R. et al. (2010). In Situ Chemical Modification of Peptide Microarrays: Application to the Study of the Antibody Responses to Methylated Antigens. In: Uttamchandani, M., Yao, S. (eds) Small Molecule Microarrays. Methods in Molecular Biology, vol 669. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-845-4_11
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DOI: https://doi.org/10.1007/978-1-60761-845-4_11
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