Microfluidic thread-based analytical devices for point-of-care detection of therapeutic antibody in blood
Microfluidic thread-based analytical devices for point-of-care detection of therapeutic antibody in blood
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DOI：
10.1016/j.snb.2021.131002
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发表时间：
2021-11-15
影响因子：
8.4
通讯作者：
Citterio, Daniel
中科院分区：
文献类型：
--
作者：
Shimazu, Riho;Tomimuro, Kosuke;Citterio, Daniel
在涉及治疗性抗体的疗法中，治疗性药物监测（TDM）具有显著提高临床疗效和实现更经济的药物使用的潜力，因为施用后从血流中的清除是患者依赖性的。由于TDM必须连续进行并持续很长一段时间，因此理想情况下应通过经济且易于进行的床旁（POC）测定来实现。在这里，我们介绍了一种新的分析方法POC TDM的临床上重要的治疗性抗体西妥昔单抗，使用微流体线程为基础的分析设备（亩TADs）作为一个平台的生物发光共振能量转移（BRET）开关传感器蛋白简单的定量抗体检测。用NaCl预处理μ M用于血浆分离，用BRET转换传感器蛋白及其底物用于西妥昔单抗检测，整合了无试剂单步测定所需的所有元素。全血应用后，生物发光发射颜色发生变化，取决于西妥昔单抗浓度。该装置设计使得仅使用2 μ L全血就能在2.5分钟内定量西妥昔单抗在治疗相关的血液浓度范围内。所获得的结果与血容量无关。这种蘑菇可以在-20摄氏度的温度下储存约一个月，这个温度与常用的冷冻机兼容。数学形态学识别（MMR）集成的图像处理技术的自动选择感兴趣的区域（ROI）的应用产生的结果与最小的错误。该方法的使用简单、快速和成本效益有望为个体患者在POC时实现治疗性抗体的TDM提供可能性。
In therapies involving therapeutic antibodies, therapeutic drug monitoring (TDM) has the potential to substantially increase clinical efficacy and to enable more economical drug use, because the clearance from the bloodstream after administration is patient dependent. Since TDM must be performed continuously and over extended time periods, it should ideally be achieved through an economical and easy to perform point-of-care (POC) assay. Here, we introduce a new analytical method for POC TDM of the clinically important therapeutic antibody cetuximab, using microfluidic thread-based analytical devices (mu TADs) as a platform for a bioluminescence resonance energy-transfer (BRET) switching sensor protein for simple quantitative antibody detection. The mu TAD is pre-treated with NaCl for blood plasma separation and a BRET switching sensor protein and its substrate for cetuximab detection, integrating all elements required for a reagent-free, single-step assay. A change in bioluminescence emission color depending on cetuximab concentrations occurs after whole blood application. The device design enables the quantification of cetuximab within the therapeutically relevant blood concentration range in 2.5 min using only 2 mu L of whole blood. The results obtained are independent of the blood volume. This mu TAD can be stored for about one month at -20 degrees C, a temperature compatible with commonly available freezers. The application of mathematical morphology recognition (MMR)-integrated image processing techniques for the automatic selection of regions of interest (ROIs) yields results with minimal error. The simplicity of use, rapidity, and cost-effectiveness of this mu TAD are expected to offer the possibility of achieving TDM of therapeutic antibodies at POC for individual patients.