Cell lysis and protein extraction in a microfluidic device with detection by a fluorogenic enzyme assay

被引:244
作者
Schilling, EA [1 ]
Kamholz, AE [1 ]
Yager, P [1 ]
机构
[1] Univ Washington, Dept Bioengn, Seattle, WA 98195 USA
关键词
D O I
10.1021/ac015640e
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A critical requirement for achieving a micro total analytical system for the analysis of cells and their constituent proteins is to integrate the lysis and fractionation steps on-chip. Here, an experimental microfluidic system integrating the lysis of bacterial cells and the extraction of a large intraceflular enzyme, beta-galactosidase, is demonstrated. The beta-galactosidase is detected and quantified using a fluorogenic enzyme assay and a numerical model. While the focus is on the lysis of typical Gram-negative bacterial cells (E. coli), the techniques described here could, in principle, be applied to a variety of different cell types.
引用
收藏
页码:1798 / 1804
页数:7
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