In this study, we describe the use of organized mesoporous titaniumoxide (TiO2) films as three-dimensional templates for protein microarrays with enhanced protein loading capacity and detection sensitivity. Multilayered mesoporous TiO2 films with high porosity and good connectivity were synthesized using a graft copolymer consisting of a poly(vinyl chloride) (PVC) backbone and poly(oxyethylene methacrylate) (POEM) side chains as a structure-directing template. The average pore size and thickness of the TiO2 films were 50-70 nm and 1.5 μm, respectively. Proteins were covalently immobilized onto mesoporous TiO2 film via 3-aminopropyltriethoxysilane (APTES), and protein loading onto TiO2 films was about four times greater than on planar glass substrates, which consequently improved the protein activity. Micropatterned mesoporous TiO 2 substrates were prepared by fabricating poly(ethylene glycol) (PEG) hydrogel microstructures on TiO2 films using photolithography. Because of nonadhesiveness of PEG hydrogel towards proteins, proteins were selectively immobilized onto surface-modified mesoporous TiO2 region, creating protein microarray. Specific binding assay between streptavidin/biotin and between PSA/anti-PSA demonstrated that the mesoporous TiO2-based protein microarrays yielded higher fluorescence signals and were more sensitive with lower detection limits than microarrays based on planar glass slides.
All Science Journal Classification (ASJC) codes
- Materials Science(all)