Department of Chemical Engineering, University of Virginia, 102 Engineers Way, Charlottesville, VA, 22901, USA.
Department of Biomedical Engineering, University of Virginia, Charlottesville, USA.
Sci Rep. 2023 Sep 13;13(1):15186. doi: 10.1038/s41598-023-41953-1.
Determination of per- and polyfluoroalkyl substances (PFAS) in drinking water at the low levels set by regulatory officials has been a major focus for sensor developing researchers. However, it is becoming more apparent that detection of these contaminants in soils, foods and consumer products is relevant and necessary at part per billion and even part per million levels. Here, a fluorescent biosensor for the rapid detection of PFOA was engineered based on human liver fatty acid binding protein (hLFABP). By conjugating circularly permuted green fluorescent protein (cp.GFP) to a split hLFABP construct, the biosensor was able to detect perfluorooctanoic acid PFOA in PBS as well as environmental water samples with LODs of 236 and 330 ppb respectively. Furthermore, E. coli cells cytosolically expressing the protein-based sensor were demonstrated to quickly detect PFOA, demonstrating feasibility of whole-cell sensing. Overall, this work demonstrates a platform technology utilizing a circularly permuted GFP and split hLFABP conjugate as a label-free optical biosensor for PFOA.
检测饮用水中监管机构设定的低水平的全氟和多氟烷基物质 (PFAS) 一直是传感器研发人员的主要关注点。然而，越来越明显的是，在土壤、食品和消费产品中检测这些污染物在十亿分之几甚至百万分之几的水平上是相关且必要的。在这里，基于人肝脂肪酸结合蛋白 (hLFABP) 设计了用于快速检测全氟辛酸 (PFOA) 的荧光生物传感器。通过将环状排列的绿色荧光蛋白 (cp.GFP) 与 hLFABP 构建体的片段连接，该生物传感器能够分别在 PBS 和环境水样中检测到全氟辛酸 (PFOA)，其检测限分别为 236 和 330 ppb。此外，还证明了细胞质中表达该蛋白的大肠杆菌细胞能够快速检测到 PFOA，证明了全细胞传感的可行性。总的来说，这项工作展示了一种利用环状排列的 GFP 和 hLFABP 片段缀合物作为无标记光学生物传感器用于检测 PFOA 的平台技术。
