Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential

Rafael Song, Stephanie Hsu, Anna L. Stevens, Jongyoon Han

Research output: Contribution to journalArticle

Abstract

Efficient sample preparation tools are the key to measuring molecular signals in a complex biological system. A novel continuous-flow isoelectric point (pI)-based sorting technique has been developed for proteins and peptides in a microfluidic chip format. It can sort biomolecules at a relatively high flow rate of up to 10 μL/min and does not require carrier ampholytes, which can create molecular backgrounds for subsequent analysis. Furthermore, the electrophoretic field required to run the pI-based sorting is generated by the diffusion of buffer ions in situ, at the liquid junction between two laminar flows within the microfluidic channel. Utilizing the diffusion potential in combination with a pH difference between the buffers, we demonstrated a separation of binary mixtures of pI markers and proteins without applying any external field. The sorting resolution and its efficiency are sufficiently high for sample preparation and could be further improved by optimizing buffers or with an additional transverse electric field. Once fully developed, it can potentially be a pI-based sample fractionation tool for proteomic analysis of complex biomolecule samples.

Original languageEnglish (US)
Pages (from-to)3528-3536
Number of pages9
JournalAnalytical Chemistry
Volume78
Issue number11
DOIs
StatePublished - Jun 1 2006

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Sorting
Microfluidics
Buffers
Biomolecules
Peptides
Ampholyte Mixtures
Proteins
Biological systems
Fractionation
Binary mixtures
Laminar flow
Electric fields
Flow rate
Ions
Liquids

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential. / Song, Rafael; Hsu, Stephanie; Stevens, Anna L.; Han, Jongyoon.

In: Analytical Chemistry, Vol. 78, No. 11, 01.06.2006, p. 3528-3536.

Research output: Contribution to journalArticle

Song, Rafael ; Hsu, Stephanie ; Stevens, Anna L. ; Han, Jongyoon. / Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential. In: Analytical Chemistry. 2006 ; Vol. 78, No. 11. pp. 3528-3536.
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