Hydrophilic modification of polystyrene with hydrophobin for time-resolved immunofluorometric assay.
Hydrophilic modification of polystyrene with hydrophobin for time-resolved immunofluorometric assay.
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DOI：
10.1016/j.bios.2010.08.059
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发表时间：
2010-11
影响因子：
12.6
通讯作者：
Zefang Wang;Yujian Huang;Shan-shan Li;Haijin Xu;M. Linder;M. Qiao
中科院分区：
文献类型：
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作者：
Zefang Wang;Yujian Huang;Shan-shan Li;Haijin Xu;M. Linder;M. Qiao
在此，我们报告了疏水蛋白膜作为固体支持物的聚苯乙烯表面上的固定抗体的时间分辨免疫荧光测定（TR-IFMA）。采用石英晶体微天平（QCM-D）、X射线光电子能谱（XPS）、水接触角（WCA）和原子力显微镜（AFM）研究了灰树花I类疏水蛋白（HGFI）对聚苯乙烯表面的亲水化修饰。通过癌胚抗原（CEA）的TR-IFMA评价HGFI修饰的聚苯乙烯的性能。QCM-D显示HGFI在pH 5下在聚苯乙烯上形成完整的单层。XPS和WCA测量表明，自组装HGFI可以使聚苯乙烯表面亲水三个月。原子力显微镜表明，一个端对抗体单层吸附在HGFI膜上，而不是在聚苯乙烯上的侧上取向的多层。CEA在5 ~ 600 ng/mL范围内线性关系良好，表明HGFI修饰的聚苯乙烯在TR-IFMA中的检测灵敏度高于未修饰的聚苯乙烯。本发明的用于改性聚苯乙烯的方法简单，不需要严格的化学处理，并且可以具有广泛的适用性，以官能化用于固定生物分子的其他载体。
Herein we reported that a hydrophobin film was used as a solid support on the polystyrene surface for immobilizing antibodies in the time-resolved immunofluorometric assay (TR-IFMA). Quartz crystal microbalance with dissipative monitoring (QCM-D), X-ray photoelectron spectroscopy (XPS) and water contact angle (WCA) measurements, as well as atomic force microscope (AFM) were used to characterize the hydrophilic modification of polystyrene surface with Class I hydrophobin isolated from Grifola frondosa (HGFI). The performance of HGFI-modified polystyrene was evaluated by TR-IFMA of carcinoembryonic antigen (CEA). QCM-D revealed that HGFI formed an intact monolayer on the polystyrene at pH 5. XPS and WCA measurements showed that self-assembling HGFI could render polystyrene surface hydrophilic for three months. AFM indicated that an end-on antibody monolayer was adsorbed on the HGFI film rather than multilayers on the polystyrene in a side-on orientation. Furthermore, a linear calibration curve (from 5 to 600ng/mL) of CEA showed HGFI-modified polystyrene had higher detection sensitivity than unmodified ones in TR-IFMA. This present method for modifying polystyrene is simple without severe chemical treatment and may have wide applicability to functionalize other supports for immobilizing biomolecules.