title: Protein nanoarray on Prolinker surface constructed by atomic force microscopy dip-pen nanolithography for analysis of protein interaction.
abstract: Protein nanoarrays are addressable ensembles of nano-scale protein domain on solid surfaces. This method can serve as a useful platform for ultraminiaturized bioanalysis. In this study, we investigated single molecular nanopatterning and molecular interaction of proteins that were immobilized on Prolinker surface of gold-coated silicon wafer by using dip-pen nanolithography (DPN) method. Contact force and humidity were optimized at 0.01 nN and 80%, respectively. The domain features of protein nanoarrays were developed at the contact time of 5 s. The optimized conditions for the nanoarray process were applied to create protein nanoarray using integrin alpha(v)beta3 and angiogenin. Constructed protein nanoarrays using integrin alpha(v)beta3 have single molecular monolayer with regular domain shape (height 15 +/- 5 nm). The changed height value due to the single molecular interaction between integrin alpha(v)beta3 and vitronectin was approximately 30 +/- 5 nm on Prolinker surface as measured with atomic force microscopy tip. Taken together, these results suggest that protein nanoarray on Prolinker surface fabricated by well-controlled DPN process can be used to analyze single molecular interaction of protein.
authors.affiliation: Protein Chip Research Center, Biotechnology Research Institute, Chungbuk National University, Cheongju, Korea.
authors.forename: Minsu
authors.initials: M
authors.lastname: Lee
authors.forename: Dong-Ku
authors.initials: DK
authors.lastname: Kang
authors.forename: Hyun-Kyu
authors.initials: HK
authors.lastname: Yang
authors.forename: Keun-Hyung
authors.initials: KH
authors.lastname: Park
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choe
authors.forename: Changsoo
authors.initials: C
authors.lastname: Kang
authors.forename: Soo-Ik
authors.initials: SI
authors.lastname: Chang
authors.forename: Moon Hi
authors.initials: MH
authors.lastname: Han
authors.forename: In-Cheol
authors.initials: IC
authors.lastname: Kang
mesh_terms: D006046:Gold; D039302:Integrin alphaVbeta3 / Q000737:chemistry* / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000379:methods*; D036103:Nanotechnology; D040081:Protein Array Analysis; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000378:metabolism; D012666:Semiconductors; D013499:Surface Properties; D019096:Vitronectin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D039302:Integrin alphaVbeta3; D019096:Vitronectin; D006046:Gold; C046366:angiogenin; D012259:Ribonuclease, Pancreatic