Micropatterned Biorecognition Surfaces on Nonbiofouling Polymer by Living Radical Photopolymerization for High Sensitivi
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1093-CC01-04
Micropatterned Biorecognition Surfaces on Nonbiofouling Polymer by Living Radical Photopolymerization for High Sensitivity Biosensing Madoka Takai1,2, James Sibarani1, and Kazuhiko Ishihara1,2 1 Department of Materials Engineering, The University of Tokyo, 7-3-1 Hongo Bunkyo-ku, Tokyo, 113-8656, Japan 2 Center for NanoBio Integration, Tokyo, 113-8656, Japan ABSTRACT Preparation of nonbiofouling surface and micropatterned biorecognition layer over nonbiofouling for highly sensitive microarray biosensors, was conducted by constructing biocompatible poly(2-methacryloyloxyethyl phosphorylcholine(MPC)) brushes on polymer substrates based on living radical polymerization using macrophotoiniferters comprised of 2ethylhexyl methacrylate (EHMA) and 4-vinylbenzyl N,N-diethyldithiocarbamate (VBDC). Protein adsorption on these modified surfaces significantly reduces compared to that on polymeric substrates, and the micropatterning consisted of poly(MPC) brush and biorecognition layer can be easily done by applying photomask with high fidelity. INTRODUCTION To create useful biomaterials for many biotechnology applications, interfaces are required that have both enhanced specific binding and reduced nonspecific binding. Thus, in applications such as biosensing, the tailoring of biointerface chemistry and the use of micro or nanofabrication technique becomes an important avenue for the production of surface with specific binding properties and minimal background interference[1]. It has already well known that cell-membrane-like surfaces based on 2-methacryloyloxyethyl phosphorylcholine (MPC) is very effective to inhibit blood coagulation on the medical devices or to prevent nonspecific biological interactions at the interface of biosensors [2, 3]. The aim of this study is to prepare micropatterned biorecognition layer on nonbiofouling surface bearing highly biocompatible poly(MPC) brushes for enhancing high signal/noise (S/N) ratio in biosensing using living radical photopolymerization based on diethyldithiocarbamate as photoiniferter (initiator, transfer, and terminator)[4]. The ideal construction of this microsensor chip devices is illustrated in figure 1. It is expected that the biomolecules conjugated on poly(MPC) brushed surface keep high activity due to the reduction of denaturation of immobilized biomolecules in the presence of the MPC unit and the chemical binding to the polymer via the oxyethylene chain[6, 7].
Figure 1. The ideal construction of micropatterned biorecoginiton layers over the nonbiofouling poly(MPC)-brushed type surface. substrate, macrophotoiniferter layer, MPC, PEG-NSC, biomolecule (protein/cell)
EXPERIMENT The macrophotoiniferters comprised of 2-ethylhexyl methacrylate (EHMA) and 4vinylbenzyl N,N-diethyldithiocarbamate (VBDC) (PEV as shown in figure 2) were synthesized with variation of VBDC content of 10% - 40% (PEV10, 20, 30, 40). Properly cleaned polyethylene terephthalate (PET) substrates were coated by dipping in 0.25 wt% of the macrophotoiniferter solutions in toluene. The macropho
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