Antibody immobilization to phospholipid polymer layer on gold substrate of quartz crystal microbalance immunosensor
Antibody immobilization to phospholipid polymer layer on gold substrate of quartz crystal microbalance immunosensor
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DOI：
10.1016/j.colsurfb.2006.11.032
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发表时间：
2007-04-01
影响因子：
5.8
通讯作者：
Ishihara, Kazuhiko
中科院分区：
文献类型：
--
作者：
Park, Jongwon;Kurosawa, Shigeru;Ishihara, Kazuhiko
采用水溶性两亲性磷脂聚合物聚[2-甲基丙烯酰氧乙基磷酰胆碱-co-甲基丙烯酸正丁酯-co-对硝基苯氧羰基聚乙二醇甲基丙烯酸酯（PMBN）]修饰免疫传感器金电极，构建人工细胞膜结构。该聚合物具有用于固定生物分子的活性酯基，并且其部分转化为用于结合到金基底的硫醇基。将部分硫醇化的PMBN吸附在石英晶体微天平（QCM）的金电极上。利用反射各向异性光谱、椭圆偏振光谱、动态接触角和X射线光电子能谱对吸附PMBN层的表面特征进行了深入研究。在具有不同硫醇化程度的几种PMBN中，可以得出结论，21.5%硫醇化的PMBN层在其外表面具有最有序的磷酸胆碱基团。蛋白质吸附实验表明，PMBN膜外侧的磷酰胆碱基团被甘氨酸取代后，对牛血清白蛋白、γ-球蛋白等蛋白质的非特异性吸附有抑制作用。此外，通过抗原-抗体结合评价，固定在PMBN表面上的抗-C-反应蛋白抗体工作良好，并且证实尽管在4 ℃下储存60天，在PMBN层上的抗体几乎不发生变性。具有良好有序的磷酸胆碱基团的抗体缀合磷脂聚合物层可以是用于生物医学诊断装置的优异的功能膜，而不会非特异性结合和降低固定化抗体的免疫活性。(C)2006 Elsevier B. V.保留所有权利。
To modify gold electrode for immunosensor to construct an artificial cell membrane structure, water-soluble amphiphilic phospholipid polymer, poly [2-methacryloyloxyehtyl phosphorylcholine-co-n-butyl methacrylate-co-p-nitrophenyloxycarbonyI poly(ethylene glycol) methacrylate (PMBN)] was applied. The polymer had active ester groups for immobilization of biomolecules and it was converted partially to thiol groups for binding to gold substrates. The partially thiolated PMBN was adsorbed on a gold electrode of quartz crystal microbalance (QCM). Surface characterization of adsorbed PMBN layers was thoroughly investigated with reflectance anisotropy spectroscopy, ellipsometry spectroscopy, dynamic contact angle and X-ray photoelectron spectroscopy measurements. Among several PMBN, having different degree of thiolation, it was concluded that 21.5% thiolated PMBN layer had the most well-ordered phosphorylcholine groups in its outer surface. The proteins adsorption test revealed that the phosphorylcholine group on the outer side of PMBN layers, which was substituted their active ester groups by glycine, showed suppress the non-specific adsorption of proteins, such as bovine serum albumin and gamma-globulin. Also, through antigen-antibody binding evaluation, the anti-C-reactive protein antibody immobilized on the PMBN surface worked well and it was confirmed that denaturation of the antibody on the PMBN layers was hardly occurred in spite of 60 days storage at 4 degrees C. The antibody conjugated phospholipid polymer layer with well-ordered phosphorylcholine group could be outstanding functional membrane for biomedical diagnostic devices without non-specific binding and reduction of immunologic activity of immobilized antibody. (C) 2006 Elsevier B.V. All rights reserved.