Institut National de la Recherche Scientifique, Énergie, Matériaux et Télécommunications (INRS-EMT), Varennes, Quebec, Canada J3X 1S2.
Biosens Bioelectron. 2013 Mar 15;41:335-41. doi: 10.1016/j.bios.2012.08.049. Epub 2012 Sep 7.
A simple sensing assay was established for label-free detection of HIV-1 protease. HIV-1 protease peptide substrate conjugated to magnetic beads via its N-terminus is directly fixed onto the sensor gold surface through the sulphur atom of cysteine. Surface plasmon resonance (SPR) was used to study the peptide substrate cleavage efficiency of the protease with magnetic beads of different sizes (1 μm and 30 nm). Cyclic voltammetry and faradic impedance spectroscopy were employed in order to characterize the functionalized gold electrode. It was found that the nano-sized beads are a more efficient sensing probe for the protease. Electrochemical biosensing showed a gradual decrease in charge transfer resistance after injection of the HIV-1 protease. The experimental data established a detection limit of 10 pg/ml, as well as demonstrated a drug screening assay. This HIV-1 protease biosensor represents a new detection approach which will lead to low-cost point-of-care devices for sensitive HIV-1 diagnosis, as well as high-throughput drug screening platforms.
建立了一种简单的传感分析方法，用于无标记检测 HIV-1 蛋白酶。通过其 N 末端将 HIV-1 蛋白酶肽底物与磁珠偶联，并通过半胱氨酸的硫原子直接固定在传感器金表面上。表面等离子体共振（SPR）用于研究不同大小（1 μm 和 30 nm）的磁珠对蛋白酶的肽底物切割效率。循环伏安法和法拉第阻抗光谱用于表征功能化金电极。结果发现，纳米级磁珠是蛋白酶更有效的传感探针。电化学生物传感显示，注入 HIV-1 蛋白酶后，电荷转移电阻逐渐降低。实验数据确定了 10 pg/ml 的检测限，并进行了药物筛选试验。这种 HIV-1 蛋白酶生物传感器代表了一种新的检测方法，将为敏感的 HIV-1 诊断以及高通量药物筛选平台带来低成本的即时检测设备。
