Development of a Practical Protein-Chip Using Designed Synthetic Peptide-Arrays

• Published in: KOBUNSHI RONBUNSHU 2004/10/25, Vol.61(10), pp.523-532
• Main Authors: NOKIHARA, Kiyoshi, OHYAMA, Takafumi, USUI, Kenji, YONEMURA, Koichi, TOMIZAKI, Kin-ya, MIHARA, Hisakazu
• Format: Article
• Language: eng ; jpn
• Subjects: Arrayer; Finger Print Method; Fluorescent Labeling; High-Throughput Synthesis; Peptide Library; Protein Chip; Protein Detection
• ISSN: 0386-2186; 1881-5685
• DOI: 10.1295/koron.61.523

Novel high-throughput technologies, which can replace conventional electrophoresis and mass spectroscopic analyses, are in great demand for understanding the structure-function relationships of proteins. For a practical protein-detection system, arrays with immobilized designed synthetic peptides have been constructed. Peptides were labeled with fluorescent dyes in order to achieve high sensitivity. Two different types of prototype-arrayer have been constructed: one has a micro-dispensing system and the other a spot-printing system. Combinatorial peptide libraries, which consisted of β-loop, β-strand and α-helical peptides, were constructed by improved highly efficient solid-phase synthesis. Labeled peptides were covalently immobilized on solid supports such as precision glass plates. Various proteins were characterized with these peptide-arrays using a fluorescent scanner to give the“protein fingerprint”which is characteristic of the individual proteins. The results of the present prototype system demonstrate the practicality of protein-chips as a new generation of biochips.