Development of Quantitative Cell-Based Enzyme Assays in Microdroplets

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• 作者：Ansgar Huebner ; Luis F. Olguin ; Daniel Bratton ; Graeme Whyte ; Wilhelm T. S. Huck ; Andrew J. de Mello ; Joshua B. Edel ; Chris Abell ; Florian Hollfelder
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：May 15, 2008
• 年：2008
• 卷：80
• 期：10
• 页码：3890 - 3896
• 全文大小：912K
• 年卷期：v.80,no.10(May 15, 2008)
• ISSN：1520-6882

文摘

We describe the development of an enzyme assay inside picoliter microdroplets. The enzyme alkaline phosphatase is expressed in Escherichia coli cells and presented in the periplasm. Droplets act as discrete reactors which retain and localize any reaction product. The catalytic turnover of the substrate is measured in individual droplets by monitoring the fluorescence at several time points within the device and exhibits kinetic behavior similar to that observed in bulk solution. Studies on wild type and a mutant enzyme successfully demonstrated the feasibility of using microfluidic droplets to provide time-resolved kinetic measurements.