A novel and rapid assay for HIV-1 protease detection using magnetic bead mediation

被引:36
作者
Esseghaier, Chiheb [1 ]
Ng, Andy [1 ]
Zourob, Mohammed [1 ]
机构
[1] INRS EMT, Varennes, PQ J3X 1S2, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
HIV-1; protease; Magnetic bead-peptide; Biosensor; Impedance spectroscopy; Surface plasmon resonance; SURFACE-PLASMON RESONANCE; MOLECULAR RECOGNITION; SENSORS; STABILIZATION; INHIBITORS; KINETICS; ANTIBODY; BINDING;
D O I
10.1016/j.bios.2012.08.049
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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 mu m 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. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:335 / 341
页数:7
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