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.
Dong
He
H
Department of Surgery, Emory University, Atlanta, GA 30332, USA.
Song
Xuezheng
X
Lasanajak
Yi
Y
Cummings
Richard D
RD
Chaikof
Elliot L
EL
D000447:Aldehydes / Q000737:chemistry; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D055767:Immobilized Proteins / Q000032:analysis / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011486:Protein C / Q000737:chemistry
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D055767:Immobilized Proteins; D010455:Peptides; D011486:Protein C