Femtomole mixer for microsecond kinetic studies of protein folding

David E. Hertzog, Xavier Michalet, Marcus Jager, Xiangxu Kong, Juan G. Santiago, Shimon Weiss, Olgica Bakajin

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

We have developed a microfluidic mixer for studying protein folding and other reactions with a mixing time of 8 μs and sample consumption of femtomoles. This device enables us to access conformational changes under conditions far from equilibrium and at previously inaccessible time scales. In this paper, we discuss the design and optimization of the mixer using modeling of convective diffusion phenomena and a characterization of the mixer performance using microparticle image velocimetry, dye quenching, and Förster resonance energy-transfer (FRET) measurements of single-stranded DNA. We also demonstrate the feasibility of measuring fast protein folding kinetics using FRET with acyl-CoA binding protein.

Original languageEnglish
Pages (from-to)7169-7178
Number of pages10
JournalAnalytical Chemistry
Volume76
Issue number24
DOIs
StatePublished - 15 Dec 2004
Externally publishedYes

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