Polymer brush biointerfaces for highly sensitive biosensors that preserve the structure and function of immobilized proteins
Polymer brush biointerfaces for highly sensitive biosensors that preserve the structure and function of immobilized proteins
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DOI：
10.1016/j.snb.2015.04.056
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发表时间：
2015-09
影响因子：
8.4
通讯作者：
K. Takasu;K. Kushiro;Katsuyoshi Hayashi;Y. Iwasaki;S. Inoue;E. Tamechika;M. Takai
中科院分区：
文献类型：
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作者：
K. Takasu;K. Kushiro;Katsuyoshi Hayashi;Y. Iwasaki;S. Inoue;E. Tamechika;M. Takai
为了开发用于蛋白质组学和生物传感器系统的高灵敏度和高通量蛋白质测定系统，我们阐明了通过不同机制固定蛋白质的各种聚合物刷表面上的蛋白质结构和活性之间的关系。通过表面引发原子转移自由基聚合（SI-ATRP）合成了不同类型的聚合物刷表面（阳离子型、疏水型、共价键合型以及阳离子型和两性离子型嵌段型），并通过圆二色谱和平衡解离常数（Kd）分析研究了固定在每个表面上的蛋白质的二级结构和活性。聚合物刷表面的二级结构的稳定性和蛋白质的活性之间有很强的相关性。有趣的是，固定在嵌段型聚合物表面（PMPC-block-PAEMA（PMBA））上的蛋白质显示出比其他聚合物刷表面更少的结构变化和更高的活性。这些结果表明，吸附诱导的结构变化的预防有很大的作用，在提高固定化蛋白质的活性，并强调两性离子聚合物的潜力，创造理想的生物界面来自功能，表面固定化蛋白质。这种生物界面可用于提高蛋白质微阵列和生物传感器系统的特异性和灵敏度。
In order to develop highly sensitive and high-throughput protein assay systems for proteomics and biosensor systems, we elucidated the relationship between protein structure and activity on various polymer brush surfaces that immobilize proteins through different mechanisms. Various polymer brush surfaces (cationic, hydrophobic, covalent-bonding, and block-type with the cationic and zwitterionic) were synthesized via surface-initiated atom transfer radical polymerization (SI-ATRP), and both the secondary structure and activity of proteins immobilized on each surface were investigated by circular dichroism and equilibrium dissociation constant (Kd) analysis, respectively. There was a strong correlation between the stability of the secondary structures and the activity of the proteins on the polymer brush surfaces. Interestingly, proteins immobilized on the block-type polymer surface (PMPC-block-PAEMA (PMbA)) showed less structural change and higher activity than the other polymer brush surfaces. These results suggest that the prevention of the adsorption-induced structural changes has a great role in improving the activity of immobilized proteins, and underscore the potentials of zwitterionic polymers for creating ideal biointerfaces derived from the functional, surface-immobilized proteins. Such biointerfaces can be useful for improving the specificity and sensitivity of protein microarrays and biosensor systems.