Hybrids of a genetically engineered antibody and a carbon nanotube transistor for detection of prostate cancer biomarkers.
Hybrids of a genetically engineered antibody and a carbon nanotube transistor for detection of prostate cancer biomarkers.
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DOI：
10.1021/nn300819s
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发表时间：
2012-06-26
期刊：
影响因子：
17.1
通讯作者：
Johnson AT
中科院分区：
文献类型：
--
作者：
Lerner MB;D'Souza J;Pazina T;Dailey J;Goldsmith BR;Robinson MK;Johnson AT
我们开发了一种新的检测方法骨桥蛋白（OPN），一种新的前列腺癌的生物标志物，通过连接基因工程单链可变片段（scFv）蛋白与OPN的高结合亲和力的碳纳米管场效应晶体管（NTFET）。使用重氮盐的化学官能化用于将scFv共价连接到NT-FET，如通过原子力显微镜证实的，同时保留OPN的生物结合位点的活性。电子传输测量表明，功能化的NT-FET可用于检测OPN与互补scFv蛋白的结合。在具有临床意义的机制中观察到源极-漏极电流的浓度依赖性增加，检测限约为30 fM。scFv-NT杂交装置表现出对OPN超过其他对照蛋白的选择性。这些装置在浓缩的牛血清白蛋白背景下对OPN的存在作出响应，而没有信号损失。基于这些观察结果，检测机制归因于碳纳米管侧壁上scFv蛋白质占据的缺陷位点处的散射变化。预期本文所述的官能化程序可推广到含有可接近的胺基的任何抗体，并产生适于检测fM浓度下的相应互补蛋白的生物传感器。
We developed a novel detection method for osteopontin (OPN), a new biomarker for prostate cancer, by attaching a genetically engineered single chain variable fragment (scFv) protein with high binding affinity for OPN to a carbon nanotube field-effect transistor (NTFET). Chemical functionalization using diazonium salts is used to covalently attach scFv to NT-FETs, as confirmed by atomic force microscopy, while preserving the activity of the biological binding site for OPN. Electron transport measurements indicate that functionalized NT-FET may be used to detect the binding of OPN to the complementary scFv protein. A concentration-dependent increase in the source-drain current is observed in the regime of clinical significance, with a detection limit of approximately 30 fM. The scFv-NT hybrid devices exhibit selectivity for OPN over other control proteins. These devices respond to the presence of OPN in a background of concentrated bovine serum albumin, without loss of signal. Based on these observations, the detection mechanism is attributed to changes in scattering at scFv protein-occupied defect sites on the carbon nanotube sidewall. The functionalization procedure described here is expected to be generalizable to any antibody containing an accessible amine group, and to result in biosensors appropriate for detection of corresponding complementary proteins at fM concentrations.
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