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

Content Provider	Springer Nature Link
Author	Lim, Chang Hwan Cho, Hyung Min Choo, Jaebum Neff, Silke Jungbauer, Alois Kumada, Yoichi Katoh, Shigeo Lee, Eun Kyu
Copyright Year	2008
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.
Starting Page	663
Ending Page	669
Page Count	7
File Format	PDF
ISSN	13872176
Journal	Biomedical Microdevices
Volume Number	11
Issue Number	3
e-ISSN	15728781
Language	English
Publisher	Springer US
Publisher Date	2009-01-14
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Peptide library Covalent immobilization Surface modification Array format Peptide screening Fluorescence intensity Engineering Fluid Dynamics Nanotechnology Biophysics/Biomedical Physics Biomedical Engineering
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Molecular Biology Biomedical Engineering

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