Perry Sarah L, Roberts Griffin W, Tice Joshua D, Gennis Robert B, Kenis Paul J A
Department of Chemical & Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
Cryst Growth Des. 2009 Jun 3;9(6):2566-2569. doi: 10.1021/cg900289d.
We report on a microfluidic method for the formation of aqueous/lipid mesophases to enable screening of suitable crystallization conditions of membrane proteins from a membrane-like phase in sub-20 nanoliter volumes. This integrated microfluidic chip and the employed mixing strategy address the specific challenges associated with the mixing of fluids of highly different viscosities (here a factor of 30) as well as the non-Newtonian character of the resulting mesophases. The chip requires less than 20 nL of material per condition screened whereas typically on the order of 10 μL or more is needed for a batch preparation in the present screening methods. We validated our approach with the successful crystallization of the membrane protein bacteriorhodopsin.
我们报告了一种用于形成水/脂质中间相的微流控方法，该方法能够在小于20纳升的体积中从类似膜的相中筛选膜蛋白的合适结晶条件。这种集成的微流控芯片和所采用的混合策略解决了与高粘度差异极大的流体（此处相差30倍）混合以及所得中间相的非牛顿特性相关的特定挑战。该芯片在每个筛选条件下所需的材料少于20纳升，而在当前的筛选方法中，批量制备通常需要10微升或更多。我们通过膜蛋白细菌视紫红质的成功结晶验证了我们的方法。
