Single-enzyme analysis in a droplet-based micro- and nanofluidic system

R. Arayanarakool, L. Shui, S.W.M. Kengen, A. van den Berg, J.C. Eijkel

Research output: Contribution to journalArticleAcademicpeer-review

38 Citations (Scopus)

Abstract

The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtoliter carriers, achieving high product concentrations from single-molecule encapsulation. The tiny droplets (f ~ 2.5-3 µm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. The method presented here allows to follow large numbers of individual droplets over time. The instrumental requirements are furthermore modest, since the small droplet size allows to use of standard microscope and standard Pyrex glass chips as well as the use of relatively high enzyme concentrations (nM range) for single molecule encapsulation
Original languageEnglish
Pages (from-to)1955-1962
JournalLab on a Chip
Volume13
Issue number10
DOIs
Publication statusPublished - 2013

Keywords

  • fluorescence microscopy
  • pyrococcus-furiosus
  • beta-glucosidase
  • propyl gallate
  • amplification
  • purification
  • evolution
  • kinetics
  • assays

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