A multiplexed immuno-sensor for on-line and automated monitoring of tissue culture protein biomarkers.
A multiplexed immuno-sensor for on-line and automated monitoring of tissue culture protein biomarkers.
复制标题
DOI：
10.1016/j.talanta.2020.122021
复制
发表时间：
2021-04
期刊：
影响因子：
6.1
通讯作者：
Z. Ramshani;Fei Fan;A. Wei;Miguel Romanello Joaquim;C. Gil;Matt George;M. Yoder;Donny Hanjaya-Putra-Donny-H
中科院分区：
文献类型：
--
作者：
Z. Ramshani;Fei Fan;A. Wei;Miguel Romanello Joaquim;C. Gil;Matt George;M. Yoder;Donny Hanjaya-Putra-Donny-H
对组织生长过程中培养基中分泌的多个蛋白质生物标志物水平的频繁在线和自动化监测对于组织工程和再生医学(TERM)产品的成功开发至关重要。在这里，我们提出了一种低成本、快速、可靠、可集成的基于阴离子交换膜(AEM)的多路传感平台。与黄金标准的人工ELISA检测不同，孵化/洗涤步骤针对每个目标进行了优化，并在微流控芯片中进行精确计量，以提高选择性。与每次使用都需要标准化的光学检测和不可靠的目视检测不同，AEM离子电流信号还提供了稳健性，这得益于离子选择膜的pH和离子强度控制能力，因此可以使用通用的标准曲线来校准所有运行。高度带电的二氧化硅纳米颗粒报告器增强了电信号，它还充当流体动力剪切放大器，以增强洗涤过程中的选择性。这个基于AEM的传感平台使用血管蛋白生物标记物内皮素-1(ET-1)、血管生成素(Ang)和胎盘生长因子(PlGF)进行测试。检测限和30年动态范围与ELISA法相当，但由于取消了校准和阻断步骤，分析时间显著缩短，分别为1小时和7小时。针对每个目标优化的协议使检测高度可靠，置信度超过98%。用三膜AEM传感器同时检测血管内皮细胞培养上清液40μ中ET-1、Ang和PlgF，验证了该平台的多重检测能力，并与ELISA法进行了比较。
Frequent on-line and automated monitoring of multiple protein biomarkers level secreted in the culture media during tissue growth is essential for the successful development of Tissue Engineering and Regenerative Medicine (TERM) products. Here, we present a low-cost, rapid, reliable, and integrable anion-exchange membrane-(AEM) based multiplexed sensing platform for this application. Unlike the gold-standard manual ELISA test, incubation/wash steps are optimized for each target and precisely metered in microfluidic chips to enhance selectivity. Unlike optical detection and unreliable visual detection for the ELISA test, which require standardization for every usage, the AEM ion current signal also offers robustness, endowed by the pH and ionic strength control capability of the ion-selective membrane, such that a universal standard curve can be used to calibrate all runs. The electrical signal is enhanced by highly charged silica nanoparticle reporters, which also act as hydrodynamic shear amplifiers to enhance selectivity during wash. This AEM-based sensing platform is tested with vascular protein biomarkers, Endothelin-1 (ET-1), Angiogenin (ANG) and Placental Growth Factor (PlGF). The limit of detection and three-decade dynamic range are comparable to ELISA assay but with a significantly reduced assay time of 1 h vs 7 h, due to the elimination of calibration and blocking steps. Optimized protocol for each target renders the detection highly reliable with more than 98% confidence. The multiplexed detection capability of the platform is also demonstrated by simultaneous detection of ET-1, ANG and PlGF in 40 μl of the vascular endothelial cell culture supernatants using three-membrane AEM sensor and the performance is validated against ELISA.