A novel peptide microarray for protein detection and analysis utilizing a dry peptide array system.
A novel dry peptide microarray system has been constructed that affords a practical solution for protein detection and analysis. This system is an array preparation and assay procedure under dry conditions that uses designed peptides as non-immobilized capture agents for the detection of proteins. The system has several advantages that include its portability and ease-of-use, as well as the fact that vaporization of sample solutions need not be considered. In this study, various proteins have been characterized with an alpha-helical peptide mini-library. When proteins were added to the peptide library array, the fluorescent peptides showed different fluorescent intensities depending on their sequences. The patterns of these responses could be regarded as 'protein fingerprints' (PFPs), which are sufficient to establish the identities of the target proteins. Furthermore, statistical analysis of the resulting PFPs was performed using cluster analysis. The PFPs of the proteins were clustered successfully depending on their families and binding properties. Additionally, the target protein was characterized using a nanolitre system and could be detected down to 1.2 fmol. These studies imply that the dry peptide array system is a promising tool for detecting and analyzing target proteins. The dry peptide array will play a role in development of high-throughput protein-detecting nano/micro arrays for proteomics and ligand screening studies.
Usui
Kenji
K
Department of Bioengineering and the COE21 program, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta-cho 4259 B-40, Midori-ku, Yokohama 226-8501, Japan.
Tomizaki
Kin-Ya
KY
Ohyama
Takafumi
T
Nokihara
Kiyoshi
K
Mihara
Hisakazu
H
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D005456:Fluorescent Dyes / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D036103:Nanotechnology; D010446:Peptide Fragments / Q000032:analysis*; D019151:Peptide Library; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis*; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D017433:Protein Structure, Secondary; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012996:Solutions / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared
D005456:Fluorescent Dyes; D010446:Peptide Fragments; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D012996:Solutions