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使用非晶硅离子敏感薄膜场效应晶体管检测DNA和蛋白质。

Detection of DNA and proteins using amorphous silicon ion-sensitive thin-film field effect transistors.

作者信息

Gonçalves D, Prazeres D M F, Chu V, Conde J P

机构信息

INESC Microsistemas e Nanotecnologias (INESC-MN), Rua Alves Redol 9, 1000-029 Lisbon, Portugal.

出版信息

Biosens Bioelectron. 2008 Dec 1;24(4):545-51. doi: 10.1016/j.bios.2008.05.006. Epub 2008 May 27.

Abstract

Amorphous silicon-based ion-sensitive field-effect transistors (a-Si:H ISFETs) are used for the label-free detection of biological molecules. The covalent immobilization of DNA, followed by DNA hybridization, and of the surface adsorption of oligonucleotides and proteins were detected electronically by the a-Si:H ISFET. The ISFET measurements are performed with an external Ag/AgCl microreference electrode immersed in 100mM phosphate buffer electrolyte with pH 7.0. Threshold voltage shifts in the transfer curve of the ISFETs are observed resulting from successive steps of surface chemical functionalization, covalent DNA attachment to the functionalized surface, surface blocking, and hybridization with a complementary target. The surface sensitivity achieved for DNA oligonucleotides is of the order of 1pmol/cm(2). Point-of-zero charge estimations were made for the functionalized surfaces and for the device surface after DNA immobilization and hybridization. The results show a correlation between the changes in the point-of-zero charge and the shift observed in the threshold voltage of the devices. Electronic detection of adsorbed proteins and DNA is also achieved by monitoring the shifts of the threshold voltage of the ISFETs, with a sensitivity of approximately 50nM.

摘要

非晶硅基离子敏感场效应晶体管(a-Si:H ISFETs)用于生物分子的无标记检测。通过a-Si:H ISFET以电子方式检测DNA的共价固定、随后的DNA杂交以及寡核苷酸和蛋白质的表面吸附。ISFET测量是在浸入pH 7.0的100mM磷酸盐缓冲电解质中的外部Ag/AgCl微参比电极下进行的。观察到ISFET转移曲线中的阈值电压偏移是由表面化学功能化、共价DNA附着到功能化表面、表面封闭以及与互补靶标杂交的连续步骤引起的。对于DNA寡核苷酸实现的表面灵敏度约为1pmol/cm²。对功能化表面以及DNA固定和杂交后的器件表面进行了零电荷点估计。结果表明零电荷点的变化与器件阈值电压中观察到的偏移之间存在相关性。通过监测ISFET阈值电压的偏移,还实现了对吸附蛋白质和DNA的电子检测灵敏,灵敏度约为50nM。

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