Department of Biomedical Engineering, Eindhoven University of Technology, 5612 AZ Eindhoven, The Netherlands.
Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5612 AZ Eindhoven, The Netherlands.
Anal Chem. 2023 May 23;95(20):7950-7959. doi: 10.1021/acs.analchem.3c00628. Epub 2023 May 13.
Industrial food processes are monitored to ensure that food is being produced with good quality, yield, and productivity. For developing innovative real-time monitoring and control strategies, real-time sensors are needed that can continuously report chemical and biochemical data of the manufacturing process. Here, we describe a generalizable methodology to develop affinity-based biosensors for the continuous monitoring of small molecules in industrial food processes. Phage-display antibody fragments were developed for the measurement of small molecules, as exemplified with the measurement of glycoalkaloids (GAs) in potato fruit juice. The recombinant antibodies were selected for use in a competition-based biosensor with single-molecule resolution, called biosensing by particle motion, using assay architectures with free particles as well as tethered particles. The resulting sensor measures GAs in the micromolar range, is reversible, has a measurement response time below 5 min, and enables continuous monitoring of GAs in protein-rich solutions for more than 20 h with concentration measurement errors below 15%. The demonstrated biosensor gives the perspective to enable a variety of monitoring and control strategies based on continuous measurement of small molecules in industrial food processes.
工业食品加工过程受到监控，以确保食品具有良好的质量、产量和生产力。为了开发创新的实时监测和控制策略，需要能够持续报告制造过程的化学和生化数据的实时传感器。在这里，我们描述了一种可推广的方法，用于开发基于亲和力的生物传感器，以连续监测工业食品加工过程中的小分子。噬菌体展示抗体片段被开发用于测量小分子，例如测量马铃薯果汁中的糖苷生物碱 (GA)。选择重组抗体用于基于竞争的生物传感器，该传感器具有单分子分辨率，称为基于粒子运动的生物传感，使用自由粒子和连接粒子的检测架构。所得传感器以微摩尔范围测量 GA，具有可逆性，测量响应时间低于 5 分钟，并且能够在富含蛋白质的溶液中连续监测 GA 超过 20 小时，浓度测量误差低于 15%。所展示的生物传感器为基于工业食品加工过程中小分子的连续测量来实现各种监测和控制策略提供了可能性。
