Biological signaling pathways are underpinned by protein switches that sense and respond to molecular inputs. Inspired by nature, engineered protein switches have been designed to directly transduce analyte binding into a quantitative signal in a simple, wash-free, homogeneous assay format. As such, they offer great potential to underpin point-of-need diagnostics that are needed across broad sectors to improve access, costs, and speed compared to laboratory assays. Despite this, protein switch assays are not yet in routine diagnostic use, and a number of barriers to uptake must be overcome to realize this potential. Here, we review the opportunities and challenges in engineering protein switches for rapid diagnostic tests. We evaluate how their design, comprising a recognition element, reporter, and switching mechanism, relates to performance and identify areas for improvement to guide further optimization. Recent modular switches that enable new analytes to be targeted without redesign are crucial to ensure robust and efficient development processes. The importance of translational steps toward practical implementation, including integration into a user-friendly device and thorough assay validation, is also discussed.

中文翻译：

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用于快速诊断测试的工程蛋白质开关

生物信号通路由感知和响应分子输入的蛋白质开关支撑。受大自然的启发，工程化蛋白质开关被设计为以简单、免洗、均质的测定形式直接将分析物结合转导为定量信号。因此，与实验室检测相比，它们提供了巨大的潜力来支持广泛部门所需的即时诊断，以改善获取、成本和速度。尽管如此，蛋白质开关分析尚未用于常规诊断用途，必须克服许多障碍才能实现这一潜力。在这里，我们回顾了用于快速诊断测试的工程蛋白质开关的机遇和挑战。我们评估了他们的设计，包括识别元素、报告器和切换机制，与性能相关并确定需要改进的领域以指导进一步优化。最新的模块化开关无需重新设计即可针对新的分析物进行定位，这对于确保稳健高效的开发过程至关重要。还讨论了转化步骤对实际实施的重要性，包括集成到用户友好的设备和彻底的检测验证中。