Research Article

A microfluidic reactor for rapid, low-pressure proteolysis with on-chip electrospray ionization

  1. Peter Liuni1,
  2. Tamanna Rob1,
  3. Derek J. Wilson1,2,*

Article first published online: 4 JAN 2010

DOI: 10.1002/rcm.4391

Issue

Rapid Communications in Mass Spectrometry

Rapid Communications in Mass Spectrometry

Volume 24, Issue 3, pages 315–320, 15 February 2010

Additional Information

How to Cite

Liuni, P., Rob, T. and Wilson, D. J. (2010), A microfluidic reactor for rapid, low-pressure proteolysis with on-chip electrospray ionization. Rapid Commun. Mass Spectrom., 24: 315–320. doi: 10.1002/rcm.4391

Author Information

  1. 1

    York University Chemistry Department, Toronto, ON, M3J 1P3, Canada

  2. 2

    Center for Research in Mass Spectrometry, Department of Chemistry, York University, Toronto, ON, M3J 1P3, Canada

*Department of Chemistry, York University, Toronto, ON, M3J 1P3.

Publication History

  1. Issue published online: 4 JAN 2010
  2. Article first published online: 4 JAN 2010
  3. Manuscript Accepted: 18 NOV 2009
  4. Manuscript Revised: 29 OCT 2009
  5. Manuscript Received: 7 SEP 2009

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Abstract

A microfluidic reactor that enables rapid digestion of proteins prior to on-line analysis by electrospray ionization mass spectrometry (ESI-MS) is introduced. The device incorporates a wide (1.5 cm), shallow (10 µm) reactor ‘well’ that is functionalized with pepsin-agarose, a design that facilitates low-pressure operation and high clogging resistance. Electrospray ionization is carried out directly from a short metal capillary integrated into the chip outlet. Fabrication, involving laser ablation of polymethyl methacrylate (PMMA), is exceedingly straightforward and inexpensive. High sequence coverage spectra of myoglobin (Mb), ubiquitin (Ub) and bovine serum albumin (BSA) digests were obtained after <4 s of residence time in the reactor. Stress testing showed little loss of performance over ∼2 h continuous use at high flow rates (30 µL/min). The device provides a convenient platform for a range of applications in proteomics and structural biology, i.e. to enable high-throughput workflows or to limit back-exchange in spatially resolved hydrogen/deuterium exchange (HDX) experiments. Copyright © 2010 John Wiley & Sons, Ltd.

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