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首页> 外文期刊>Crystal growth & design >A Chemical Library to Screen Protein and Protein-Ligand Crystallization Using a Versatile Microfluidic Platform
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A Chemical Library to Screen Protein and Protein-Ligand Crystallization Using a Versatile Microfluidic Platform

机译：使用通用的微流体平台筛选筛选蛋白质和蛋白质 - 配体结晶的化学文库

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摘要

Here, we describe a plug-and-play microfluidic platform, suitable for protein crystallization. The droplet factory is designed to generate hundreds of droplets as small as a few nanoliters (similar to 2 to 10 nL) for screening and optimization of crystallization conditions. Commercially available microfluidic junctions and tubing are combined to create the appropriate geometry. In addition, a "chemical library" is produced in tubing. The microfluidic geometry for a "crystallization agent-based chemical library" is validated by screening crystallization conditions of lysozyme. The microfluidic geometry for a "ligand-based chemical library" is also explored to co-crystallize the protein QR2 with a ligand, for the purposes of structure-based drug design. This platform mixes aqueous phases (containing the protein and the crystallization agent) and organic phases (containing the ligand), during the droplet generation and circulation without using any surfactant. The droplet composition is controlled by the respective flow rates of the different solutions, and checked by measuring online absorbance. The low volumes involved in the crystallization trials, the speed of execution, and the absence of a microfabrication stage make our platform a cheap, easy-to-use, and versatile tool for crystallization studies.
机译：在这里，我们描述了一种即插即用的微流体平台，适用于蛋白质结晶。液滴工厂设计用于产生数百只液滴，作为少数纳米（类似于2至10 NL），用于筛选和优化结晶条件。商业上可获得的微流体连接和管道组合以产生适当的几何形状。此外，在管道中产生“化学文库”。通过筛选溶菌酶的结晶条件来验证“结晶剂基化学文库”的微流体几何形状。对于结构的药物设计的目的，还探讨了“配体基化学文库”的微流体几何形状以将蛋白质QR2与配体共结晶。该平台在液滴产生和循环期间混合水相（含有蛋白质和结晶剂）和有机相（含有配体）而不使用任何表面活性剂。液滴组合物由不同溶液的各自流速控制，并通过测量在线吸光度来检查。涉及结晶试验的低体积，执行速度和微制造阶段的缺失使我们的平台成为结晶研究的廉价，易用和多功能的工具。

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  • 来源
    《Crystal growth & design》 |2018年第9期|共8页
  • 作者

    Gerard Charline J. J.; Ferry Gilles; Vuillard Laurent M.; Boutin Jean A.; Ferte Nathalie; Grossier Romain; Candoni Nadine; Veesler Stephane;

  • 作者单位

    Aix Marseille Univ CINaM CNRS Campus Luminy Case 913 F-13288 Marseille 09 France;

    Inst Rech Serv 125 Chemin Ronde F-78290 Croissy Sur Seine France;

    Inst Rech Serv 125 Chemin Ronde F-78290 Croissy Sur Seine France;

    Inst Rech Serv 125 Chemin Ronde F-78290 Croissy Sur Seine France;

    Aix Marseille Univ CINaM CNRS Campus Luminy Case 913 F-13288 Marseille 09 France;

    Aix Marseille Univ CINaM CNRS Campus Luminy Case 913 F-13288 Marseille 09 France;

    Aix Marseille Univ CINaM CNRS Campus Luminy Case 913 F-13288 Marseille 09 France;

    Aix Marseille Univ CINaM CNRS Campus Luminy Case 913 F-13288 Marseille 09 France;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 晶体学;
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