The specificity of biological systems makes it possible to develop biosensors targeting specific metabolites, toxins, and pollutants in complex medical or environmental samples without interference from structurally similar compounds. For the last two decades, great efforts have been devoted to creating proteins or nucleic acids with novel properties through synthetic biology strategies. Beyond augmenting biocatalytic activity, expanding target substrate scopes, and enhancing enzymes' enantioselectivity and stability, an increasing research area is the enhancement of molecular specificity for genetically encoded biosensors. Here, we summarize recent advances in the development of highly specific biosensor systems and their essential applications. First, we describe the rational design principles required to create libraries containing potential mutants with less promiscuity or better specificity. Next, we review the emerging high-throughput screening techniques to engineer biosensing specificity for the desired target. Finally, we examine the computer-aided evaluation and prediction methods to facilitate the construction of ligand-specific biosensors.

生物系统的特异性使得开发针对复杂医学或环境样品中特定代谢物、毒素和污染物的生物传感器成为可能，而不受结构相似化合物的干扰。在过去的二十年中，人们致力于通过合成生物学策略创造具有新特性的蛋白质或核酸。除了增强生物催化活性、扩大目标底物范围以及增强酶的对映选择性和稳定性之外，越来越多的研究领域是增强基因编码生物传感器的分子特异性。在这里，我们总结了高度特异性生物传感器系统及其基本应用开发的最新进展。首先，我们描述了创建包含杂乱程度较低或特异性较高的潜在突变体的文库所需的合理设计原则。接下来，我们回顾新兴的高通量筛选技术，以设计所需目标的生物传感特异性。最后，我们研究了计算机辅助评估和预测方法，以促进配体特异性生物传感器的构建。