Spot arrays on modified glass surfaces for efficient SPOT synthesis and on‐chip bioassay of peptides

To make SPOT synthesis of peptides and their assays on glass surfaces more convenient, a simple method for making spot arrays on a slide glass was designed through patterning with a photoresist and perfluorination followed by amination with various silane compounds and polymers. With these spot‐arrayed glass surfaces, we could measure the coupling completion of each Fmoc amino acid on the glass surface by direct fluorescence analysis after fluorescence‐labeling the amino groups on the surface of each spot. Then we synthesized several types of decapeptides and HPQ‐pentapeptides on the spot‐arrayed glasses and identified the optimal surface condition for stepwise peptide coupling and on‐chip bioassay. After optimizing the surface conditions, we synthesized a model library of biotin‐Gly‐Ala‐P1‐Gly (P1: one of 19 amino acids) and successfully replicated the well‐known α‐chymotrypsin subsite specificities through Cy5‐streptavidin binding to the remaining biotin on the surface after the enzymatic digestion. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd.

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