Cell lysis and protein extraction in a microfluidic device with detection by a fluorogenic enzyme assay.
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 intracellular 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.
Schilling
Eric A
EA
Department of Bioengineering, University of Washington, Seattle 98195, USA.
Kamholz
Andrew Evan
AE
Yager
Paul
P
D001426:Bacterial Proteins / Q000032:analysis*; D004926:Escherichia coli / Q000201:enzymology*; D008954:Models, Biological; D001616:beta-Galactosidase / Q000032:analysis*
D001426:Bacterial Proteins; D001616:beta-Galactosidase