Specific adsorption of histidine-tagged proteins on silica surfaces modified with Ni2+/NTA-derivatized poly(ethylene glycol)
- Authors
- Kang, Eunah; Park, Jin-Won; McClellan, Scott J.; Kim, Jong-Mok; Holland, David P.; Lee, Gil U.; Franses, Elias I.; Park, Kinam; Thompson, David H.
- Issue Date
- May-2007
- Publisher
- AMER CHEMICAL SOC
- Citation
- LANGMUIR, v.23, no.11, pp 6281 - 6288
- Pages
- 8
- Journal Title
- LANGMUIR
- Volume
- 23
- Number
- 11
- Start Page
- 6281
- End Page
- 6288
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/48022
- DOI
- 10.1021/la063719e
- ISSN
- 0743-7463
1520-5827
- Abstract
- Silica surfaces modified with nitrilotriacetic acid (NTA)-polyethylene glycol (PEG) derivatives were used to immobilize hexahistidine-tagged green fluorescent protein (His(6)-GFP), biotin/streptavidin-AlexaFluor555 (His(6)-biotin/SA-AF), and gramicidin A-containing vesicles (His(6)-gA). Three types of surface-reactive PEG derivatives-NTA-PEG3400-Si(OMe)(3), NTA-PEG3400-vinylsulfone, and mPEG5000-Si(OMe)(3) (control)-were grafted onto silica and tested for their ability to capture His(6)-tag species via His(6)/Ni2+/NTA chelation. The composition and thicknesses of the PEG-modified surfaces were characterized using X-ray photoelectron spectroscopy, contact angle, and ellipsometry. Protein capture efficiencies of the NTA-PEG-grafted surfaces were evaluated by measuring fluorescence intensities of these surfaces after exposure to His(6)-tag species. XPS and ellipsometry data indicate that surface adsorption occurs via specific interactions between the His(6)-tag and the Ni2+/NTA-PEG-grafted surface. Protein immobilization was most effective for NTA-PEG3400-Si(OMe)(3)-modified surfaces, with maximal areal densities achieved at 45 pmol/cm(2) for His(6)-GFP and 95 fmol/cm(2) for His(6)-biotin/SA-AF. Lipid vesicles containing His(6)-gA in a 1:375 gA/lipid ratio could also be immobilized on Ni2+/NTA-PEG3400-Si(OMe)(3)-modified surfaces at 0.5 mM total lipid. Our results suggest that NTA-PEG-Si(OMe)(3) conjugates may be useful tools for immobilizing His(6)-tag proteins on solid surfaces to produce protein-functionalized surfaces.
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - College of Engineering > School of Chemical Engineering and Material Science > 1. Journal Articles
![qrcode](https://api.qrserver.com/v1/create-qr-code/?size=55x55&data=https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/48022)
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.