聚(甲基丙烯酸-2-羟乙酯)改性金表面用于构建生物检测基材

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采用电子活化再生原子转移自由基聚合(AGET ATRP)的方法将聚(甲基丙烯酸-2-羟乙酯)(PHEMA)接枝在金表面,对经修饰的金表面的生物惰性做了系统的研究,并利用PHEMA的羟基末端固定生物素(biotin)分子,以biotin对抗生物素蛋白(avidin)的识别为模型,研究了不同厚度的PHEMA对结合avidin的影响,以及该表面作为生物检测基材的可行性.生物惰性研究表明,PHEMA修饰的金表面不但能够有效的排斥纤维蛋白原(Fg)、人血清白蛋白(HSA)和溶菌酶(Lys)的非特异性吸附,还能够抑制3种细胞(L02、L929和EC)的黏附,是一种良好的抗污表面.通过控制聚合时间制备了不同厚度的PHEMA-biotin修饰的表面,同位素125I标记HSA吸附结果表明这几种表面均能够有效排斥非特异性蛋白质吸附,特异性FITC-avidin吸附结果表明,厚度较小时(16 nm)由于荧光淬灭而难以检测到荧光信号,厚度在16 nm和49 nm之间,荧光信号随厚度增加而增强,通过比较信噪比,认为厚度在49 nm以上时比较理想.该表面在应用于QCM与荧光检测中均表现出良好的检测性能. Poly (2-hydroxyethyl methacrylate) (PHEMA) was grafted onto the gold surface using a method of electron-activated regenerative atom transfer radical polymerization (AGET ATRP) to systematically modify the biological inertness of the modified gold surface Using PHEMA as a biotin molecule, biotin was used as a model to identify avidin. The effects of PHEMA with different thickness on the binding of avidin were studied, and the surface was used as a bioassay substrate Bio-inert studies have shown that, PHEMA modified gold surface not only can effectively exclude the non-specific adsorption of fibrinogen (Fg), human serum albumin (HSA) and lysozyme (Lys), but also can inhibit the three kinds of cells (L02, L929 and EC) was a good anti-fouling surface.The PHEMA-biotin modified surface with different thickness was prepared by controlling the polymerization time.The results of isotope 125I-labeled HSA adsorption showed that these two surfaces could effectively repel Non-specific protein adsorption and specific FITC-avidin adsorption results showed that the fluorescence signal was hard to detect due to fluorescence quenching when the thickness was small (16 nm), the thickness was between 16 nm and 49 nm, and the fluorescence signal increased with increasing thickness In addition, by comparing the signal to noise ratio, it is considered that the thickness is more than 49 nm, and the surface shows good detection performance when applied to QCM and fluorescence detection.
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