Key Laboratory for Thin Film and Microfahrication of Ministry of Education, Research Institute of Micro/Nano Science and Technology, Shanghai JiaoTong University, Shanghai, China.
School of Physics and Electronic Engineering, Xinyang Normal University, Xinyang, China.
PLoS One. 2018 Mar 30;13(3):e0194631. doi: 10.1371/journal.pone.0194631. eCollection 2018.
A sensitive and innovative assay system based on a micro-MEMS-fluxgate sensor and immunomagnetic beads-labels was developed for the rapid analysis of C-reactive proteins (CRP). The fluxgate sensor presented in this study was fabricated through standard micro-electro-mechanical system technology. A multi-loop magnetic core made of Fe-based amorphous ribbon was employed as the sensing element, and 3-D solenoid copper coils were used to control the sensing core. Antibody-conjugated immunomagnetic microbeads were strategically utilized as signal tags to label the CRP via the specific conjugation of CRP to polyclonal CRP antibodies. Separate Au film substrates were applied as immunoplatforms to immobilize CRP-beads labels through classical sandwich assays. Detection and quantification of the CRP at different concentrations were implemented by detecting the stray field of CRP labeled magnetic beads using the newly-developed micro-fluxgate sensor. The resulting system exhibited the required sensitivity, stability, reproducibility, and selectivity. A detection limit as low as 0.002 μg/mL CRP with a linearity range from 0.002 μg/mL to 10 μg/mL was achieved, and this suggested that the proposed biosystem possesses high sensitivity. In addition to the extremely low detection limit, the proposed method can be easily manipulated and possesses a quick response time. The response time of our sensor was less than 5 s, and the entire detection period for CRP analysis can be completed in less than 30 min using the current method. Given the detection performance and other advantages such as miniaturization, excellent stability and specificity, the proposed biosensor can be considered as a potential candidate for the rapid analysis of CRP, especially for point-of-care platforms.
一种基于微机电系统-磁通门传感器和免疫磁珠标记的灵敏且创新的检测系统被开发用于快速分析 C 反应蛋白(CRP)。本研究中提出的磁通门传感器是通过标准微机电系统技术制造的。采用由 Fe 基非晶带材制成的多环磁芯作为传感元件,并使用 3-D 螺线管铜线圈来控制传感核心。抗体偶联的免疫磁珠被策略性地用作信号标记物,通过 CRP 与多克隆 CRP 抗体的特异性结合来标记 CRP。单独的 Au 薄膜基底被用作免疫平台,通过经典的三明治测定法来固定 CRP 珠标记物。通过使用新开发的微磁通门传感器检测 CRP 标记磁珠的 stray 场来实现对不同浓度 CRP 的检测和定量。所得到的系统表现出所需的灵敏度、稳定性、重现性和选择性。实现了低至 0.002μg/mL CRP 的检测限,线性范围从 0.002μg/mL 到 10μg/mL,这表明所提出的生物系统具有高灵敏度。除了极低的检测限之外,该方法还易于操作且具有快速的响应时间。我们的传感器的响应时间小于 5s,并且使用当前方法,CRP 分析的整个检测周期可以在不到 30 分钟内完成。鉴于检测性能以及其他优势,如小型化、优异的稳定性和特异性,所提出的生物传感器可被视为 CRP 快速分析的潜在候选者,特别是用于即时检测平台。