Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry

Rafael Song, Michael Chan, Chris Cello, Steven R. Tannenbaum, John S. Wishnok, Jongyoon Han

Research output: Contribution to journalArticle

Abstract

In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pi range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a highthroughput sample fractionation tool for targeted proteomics using LC/MS.

Original languageEnglish (US)
Pages (from-to)2317-2325
Number of pages9
JournalAnalytical Chemistry
Volume82
Issue number6
DOIs
StatePublished - Mar 15 2010

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Electrophoresis
Sorting
Mass spectrometry
Peptides
Buffers
Proteins
Fabrication
Substrates
Fractionation
Titration
Printing
Salts
Plasmas
Glass

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry. / Song, Rafael; Chan, Michael; Cello, Chris; Tannenbaum, Steven R.; Wishnok, John S.; Han, Jongyoon.

In: Analytical Chemistry, Vol. 82, No. 6, 15.03.2010, p. 2317-2325.

Research output: Contribution to journalArticle

Song, Rafael ; Chan, Michael ; Cello, Chris ; Tannenbaum, Steven R. ; Wishnok, John S. ; Han, Jongyoon. / Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry. In: Analytical Chemistry. 2010 ; Vol. 82, No. 6. pp. 2317-2325.
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