Nano Archive

Fluorescence-based peptide screening using ligand peptides directly conjugated to a thiolated glass surface

Lim, Chang Hwan and Cho, Hyung Min and Choo, Jaebum and Neff, Silke and Jungbauer, Alois and Kumada, Yoichi and Katoh, Shigeo and Lee, Eun Kyu (2009) Fluorescence-based peptide screening using ligand peptides directly conjugated to a thiolated glass surface. Biomedical Microdevices, 11 (3). pp. 663-669.

[img]
Preview
PDF
233Kb

Abstract

Functional peptides from peptide libraries are frequently screened using an array format. We report here results of a feasibility study of fluorescence-based peptide screening using an array format on surface-modified glass. The surface of an amine-coated glass slide was modified to contain thiol groups by iminothiolane treatment. The ε-amine of the C-terminal lysine from a ligand peptide was iodinated and then spotted onto the thiolated glass surface to covalently conjugate the ligand peptide to the surface via a thioether bond. This covalent immobilization allowed the ligand peptides to withstand washing steps by tightly adhering to the glass surface and confining their subsequent binding reactions within a spotted area. Two representative peptides were used as the ligand peptides; a ‘target’ (positive) heptapeptide that could specifically bind to trypsin, and a ‘control’ (negative) hexapeptide that had no binding affinity with trypsin. When fluorescein isothiocyanate-labeled trypsin was reacted with the ligand peptides, the target peptide demonstrated distinctively higher (ca. 8.7-fold) fluorescence intensity that was easily differentiated from the control peptide by a fluorescence scanner. A separate experiment using a quartz crystal microbalance confirmed that the difference in binding mass (ca. 9.1-fold) was very close to that seen in fluorescence intensity. These results suggested a quantitative, 1:1 correlation between mass and fluorescence signals. Furthermore, a smaller spot volume and a higher ligand peptide concentration resulted in higher fluorescence signal intensity. This study provides information on the potential for using fluorescence-based screening of functional peptides on a glass array format.

Item Type:Article
Subjects:Physical Science > Nano objects
Material Science > Nanostructured materials
Material Science > Nanochemistry
ID Code:5096
Deposited By:CSMNT
Deposited On:25 Aug 2010 11:28
Last Modified:25 Aug 2010 11:28

Repository Staff Only: item control page