Michielsen Claire M S, Lin Yu-Ting, Yan Junhong, de Jong Arthur M, Prins Menno W J
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5612 AE, the Netherlands.
Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5612 AE, the Netherlands.
ACS Sens. 2025 Apr 25;10(4):2895-2905. doi: 10.1021/acssensors.4c03637. Epub 2025 Mar 24.
Sensing technologies for the continuous monitoring of protein concentrations are important for understanding time-dependent behaviors of biological systems and for controlling bioprocesses. We present a continuous sensing methodology based on tethered particle motion (t-BPM) that utilizes fast-dissociating antibody fragments (Fabs) for continuous protein monitoring. A competition-based t-BPM sensor was developed and characterized utilizing custom-made Fabs. The sensing concept was demonstrated for lactoferrin, an 80 kDa iron-binding glycoprotein that is part of the innate immune response. Thirteen Fabs were compared using free particle motion sensing as well as surface plasmon resonance, of which six Fabs showed rapid association and dissociation. The integration of the Fabs into the t-BPM sensor enabled nanomolar lactoferrin detection in both buffer solutions and milk matrices over tens of hours. This work demonstrates how continuous protein sensing can be realized using fast-dissociating antibodies in a competitive sensor format.
用于连续监测蛋白质浓度的传感技术对于理解生物系统的时间依赖性行为以及控制生物过程至关重要。我们提出了一种基于 tethered particle motion (t-BPM) 的连续传感方法，该方法利用快速解离抗体片段 (Fabs) 进行连续蛋白质监测。利用定制的 Fabs 开发并表征了一种基于竞争的 t-BPM 传感器。针对乳铁蛋白（一种 80 kDa 的铁结合糖蛋白，是先天免疫反应的一部分）展示了传感概念。使用自由粒子运动传感以及表面等离子体共振对 13 种 Fabs 进行了比较，其中 6 种 Fabs 显示出快速结合和解离。将 Fabs 整合到 t-BPM 传感器中能够在缓冲溶液和牛奶基质中长达数十小时内检测到纳摩尔级的乳铁蛋白。这项工作展示了如何在竞争传感器形式中使用快速解离抗体实现连续蛋白质传感。
