Facile construction of fluorescent peptide microarrays: One-step fluorescent derivatization of sub-microscale peptide aldehydes for selective terminal immobilization.

In this note, we demonstrate the utility of bifunctional fluorescent linkers to facilitate the construction of peptide microarrays with either an N- or a C-terminal alkylamine for directionally preferred peptide immobilization. Significantly, these small tags facilitate high-performance liquid chromatography (HPLC) profiling while limiting interference with antigen-antibody interactions after peptide immobilization. In a model peptide-antibody binding assay, a sequence-dependent orientation effect of antibody binding to a series of peptide ligands was demonstrated. This approach provides a strategy that can be applied to a variety of peptide microarray-based detection systems.