McSweeney Megan A, Patterson Alexandra T, Loeffler Kathryn, de Larrea Regina Cuellar Lelo, McNerney Monica P, Kane Ravi S, Styczynski Mark P
School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, United States.
bioRxiv. 2024 Jul 19:2024.07.19.604303. doi: 10.1101/2024.07.19.604303.
Conventional laboratory protein detection techniques are not suitable for point-of-care (POC) use because they require expensive equipment and laborious protocols, and existing POC assays suffer from long development timescales. Here, we describe a modular cell-free biosensing platform for generalizable protein detection that we call TLISA (7 RNA polymerase-inked mmunoensing ssay), designed for extreme flexibility and equipment-free use. TLISA uses a split T7 RNA polymerase fused to affinity domains against a protein. The target antigen drives polymerase reassembly, inducing reporter expression. We characterize the platform, then demonstrate its modularity by using 16 affinity domains against four different antigens with minimal protocol optimization. We show TLISA is suitable for POC use by sensing human biomarkers in serum and saliva with a colorimetric readout within one hour and by demonstrating functionality after lyophilization. Altogether, this technology could have potentially revolutionary impacts, enabling truly rapid, reconfigurable, equipment-free detection of virtually any protein.
传统的实验室蛋白质检测技术不适用于即时检测（POC），因为它们需要昂贵的设备和繁琐的操作流程，而且现有的即时检测方法开发时间长。在此，我们描述了一种用于通用蛋白质检测的模块化无细胞生物传感平台，我们称之为TLISA（7种RNA聚合酶连接免疫传感分析），其设计具有极高的灵活性且无需设备。TLISA使用与针对蛋白质的亲和结构域融合的分裂T7 RNA聚合酶。目标抗原驱动聚合酶重新组装，诱导报告基因表达。我们对该平台进行了表征，然后通过使用针对四种不同抗原的16个亲和结构域，在最小化方案优化的情况下展示了其模块化。我们通过在一小时内以比色法读出检测血清和唾液中的人类生物标志物，并通过展示冻干后的功能，表明TLISA适用于即时检测。总之，这项技术可能会产生潜在的革命性影响，能够实现对几乎任何蛋白质的真正快速、可重新配置、无需设备的检测。
