使用纳升尺寸液滴在微流控芯片上筛选蛋白质结晶条件。

Screening of protein crystallization conditions on a microfluidic chip using nanoliter-size droplets.

作者信息

Zheng Bo, Roach L Spencer, Ismagilov Rustem F

机构信息

Department of Chemistry, The University of Chicago, 5735 South Ellis Avenue, Chicago, Illinois 60637, USA.

出版信息

J Am Chem Soc. 2003 Sep 17;125(37):11170-1. doi: 10.1021/ja037166v.

DOI:10.1021/ja037166v

PMID:16220918

Abstract

Protein crystallization is a major bottleneck in determining tertiary protein structures from genomic sequence data. This paper describes a microfluidic system for screening hundreds of protein crystallization conditions using less than 4 nL of protein solution for each crystallization droplet. The droplets are formed by mixing protein, precipitant, and additive stock solutions in variable ratios in a flow of water-immiscible fluids inside microchannels. Each droplet represents a discrete trial testing different conditions. The system has been validated by crystallization of several water-soluble proteins.

摘要

蛋白质结晶是从基因组序列数据确定蛋白质三级结构的主要瓶颈。本文描述了一种微流控系统，该系统可使用少于4纳升的蛋白质溶液来筛选数百种蛋白质结晶条件，每个结晶微滴均如此。微滴是通过在微通道内不与水混溶的流体流中以可变比例混合蛋白质、沉淀剂和添加剂储备溶液而形成的。每个微滴代表一个测试不同条件的独立试验。该系统已通过几种水溶性蛋白质的结晶得到验证。