Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
Curr Opin Struct Biol. 2015 Aug;33:42-51. doi: 10.1016/j.sbi.2015.06.001. Epub 2015 Sep 3.
The success of a directed evolution approach to creating custom-made enzymes relies in no small part on screening as many clones as possible. The miniaturisation of assays into pico to femtoliter compartments (emulsion droplets, vesicles or gel-shell beads) makes directed evolution campaigns practically more straightforward than current large scale industrial screening that requires liquid handling equipment and much manpower. Several recent experimental formats have established protocols to screen more than 10 million compartments per day, representing unprecedented throughput at low cost. This review introduces the emerging approaches towards making biomimetic man-made compartments that are poised to be adapted by a wider circle of researchers. In addition to cost and time saving, control of selection pressures and conditions, the quantitative readout that reports on every library members and the ability to develop strategies based on these data will increase the degrees of freedom in designing and testing strategies for directed evolution experiments.
定向进化方法创建定制酶的成功在很大程度上依赖于尽可能多地筛选克隆。将测定法微型化为皮升到飞升体积（乳液液滴、囊泡或凝胶壳珠），使得定向进化实验比当前需要液体处理设备和大量人力的大规模工业筛选更加简单。最近的几种实验方案已经建立了每天筛选超过 1000 万个隔室的方案，以低成本实现了前所未有的通量。本综述介绍了正在出现的仿生人造隔室的方法，这些方法有望被更广泛的研究人员所采用。除了节省成本和时间外，还可以控制选择压力和条件、进行定量读数以报告每个文库成员，并能够根据这些数据制定策略，这将增加设计和测试定向进化实验策略的自由度。
