McLeod Jennifer, Park Chankyu, Cunningham Alexandra, O'Donnell Lynne, Brown R Stephen, Kelly Fiona, She Zhe
Department of Chemistry, Queen's University, Kingston, ON K7L 3N6, Canada.
Analyst. 2020 Sep 14;145(18):6024-6031. doi: 10.1039/d0an01050b.
The biorecognition ability of hybridized toll-like receptors (TLRs) 2 and 6 proteins on electrode surfaces has been studied. TLR biosensors have been designed to be non-specific to particular bacterial strains but rather to provide broad spectrum detection of cells and toxins containing relevant pathogen-associated molecular patterns (PAMPs). Our electrochemical TLR2/6 biosensors demonstrated selective detection towards Gram-positive bacterial whole-cells and a synthetic diacylated lipopeptide (Pam2CSK4), a PAMP. Responses towards Bacillus licheniformis (B. licheniformis) and Enterococcus hirae (E. hirae) were obtained. The biosensor was able to differentiate signals between B. licheniformis and a Gram-negative bacterial cell (control) as low as 100 CFU mL-1. One challenge in developing protein-based biosensors is to improve the shelf-life of the biosensor chips and preserve the detection activity of the protein molecules, therefore we did our first exploration into storage conditions. The activity of stored biosensors was found to be strongly dependent on storage medium, and that effective 'shelf-life' was obtained makes an important step towards creating robust sensors for real-life applications.
研究了电极表面杂交的Toll样受体（TLR）2和6蛋白的生物识别能力。TLR生物传感器被设计为对特定细菌菌株无特异性，而是用于对含有相关病原体相关分子模式（PAMP）的细胞和毒素进行广谱检测。我们的电化学TLR2/6生物传感器对革兰氏阳性细菌全细胞和一种合成的二酰化脂肽（Pam2CSK4，一种PAMP）表现出选择性检测。获得了对地衣芽孢杆菌（B.licheniformis）和平肠球菌（E.hirae）的响应。该生物传感器能够区分地衣芽孢杆菌和革兰氏阴性细菌细胞（对照）之间低至100 CFU mL-1的信号。开发基于蛋白质的生物传感器的一个挑战是提高生物传感器芯片的保质期并保持蛋白质分子的检测活性，因此我们首次探索了储存条件。发现储存的生物传感器的活性强烈依赖于储存介质，并且获得有效的“保质期”是朝着为实际应用创建强大传感器迈出的重要一步。
