A centrifugal microfluidic device for screening protein crystallization conditions by vapor diffusion

A centrifugal microfluidic device for screening protein crystallization conditions by vapor diffusion
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一种通过蒸汽扩散筛选蛋白质结晶条件的离心微流体装置

DOI:
10.1016/j.snb.2015.04.105
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发表时间:
2015-11
期刊:
Sensors and Actuators B: Chemical
影响因子:
--
通讯作者:
Zhao, Jianlong
Zhao, Jianlong
中科院分区:
其他
文献类型:
--
作者:
Hu, Xiaojian;Li, Gang;Jin, Qinghui;Zhao, Jianlong

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本文描述了一种基于离心的微流控装置,用于通过气相扩散进行纳升级蛋白质结晶试验。该装置结合毛细管效应和离心力,自发地将蛋白质样品和沉淀物定量和分配到24个反应室中进行结晶筛选。此外,该装置中的两级毛细管截止阀和蒸气扩散室的设计允许蛋白质液滴和储存的沉淀剂之间的蒸气输送,完全等同于在常规蒸气扩散技术中观察到的,其缓慢地使蛋白质液滴进入过饱和状态以成核和生长蛋白质晶体。我们证明了这个设备的力量,通过测试它的蛋白质结晶条件的快速筛选。与传统的悬滴法相比,该装置可以识别更多的结晶条件,提供更宽的过饱和度范围,而蛋白质样品的使用量减少了两个数量级。这种基于离心的微流体装置为各个实验室提供了一种简单且低成本的工具,以快速筛选初始蛋白质结晶条件。
This paper describes a centrifuge-based microfluidic device for performing nanoliter-scale protein crystallization trials by vapor diffusion. By combining the capillary effect and centrifugal force, this device spontaneously and parallelly meters and dispenses protein samples and precipitants into 24 reaction chambers for crystallization screening. Furthermore, the design of two-level capillary stop valves and vapor-diffusion chambers in this device allows vapor transport between the protein droplet and the stored precipitant, perfectly equivalent to that observed in conventional vapor diffusion techniques, which slowly brings the protein droplet into a supersaturation state to nucleate and grow protein crystals. We demonstrate the power of this device by testing it for rapid screening of protein crystallization conditions. Compared with the conventional hanging-drop technique, this device can identify more crystallization conditions and provide a wider range of supersaturation rates, while using two orders of magnitude less protein sample. This centrifuge-based microfluidic device provides a simple and low-cost tool for individual laboratories to rapidly screen initial protein crystallization conditions.
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