Monolithic porous polymer stationary phases in polyimide chips for the fast high-performance liquid chromatography separation of proteins and peptides

Pavel A. Levkin, Sebastiaan Eeltink, Thomas R. Stratton, Reid Brennen, Karla Robotti, Hongfeng Yin, Kevin Killeen, Frantisek Svec, Jean M.J. Fréchet*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

107 Scopus citations

Abstract

Poly(lauryl methacrylate-co-ethylene dimethacrylate) and poly(styrene-co-divinylbenzene) stationary phases in monolithic format have been prepared by thermally initiated free radical polymerization within polyimide chips featuring channels having a cross-section of 200 μm × 200 μm and a length of 6.8 cm. These chips were then used for the separation of a mixture of proteins including ribonuclease A, myoglobin, cytochrome c, and ovalbumin, as well as peptides. The separations were monitored by UV adsorption. Both the monolithic phases based on methacrylate and on styrene chemistries enabled the rapid baseline separation of most of the test mixtures. Best performance was achieved with the styrenic monolith leading to fast baseline separation of all four proteins in less than 2.5 min. The in situ monolith preparation process affords microfluidic devices exhibiting good batch-to-batch and injection-to-injection repeatability.

Original languageEnglish (US)
Pages (from-to)55-61
Number of pages7
JournalJournal of Chromatography A
Volume1200
Issue number1
DOIs
StatePublished - Jul 18 2008
Externally publishedYes

Bibliographical note

Funding Information:
Support of this research by a grant of the National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health (EB-006133) is gratefully acknowledged. Measurement work at the Molecular Foundry was supported by the Director, Office of Science, Office of Basic Energy Sciences, Division of Materials Sciences and Engineering, of the US Department of Energy under Contract No. DE-AC02-05CH11231. T.S. is grateful to the US Department of Homeland Security for a scholarship. Agilent Technologies, Inc. is acknowledged for kindly supplying chip-related instrumentation and Debbie Ritchey at Agilent Laboratories is thanked for fabricating the prototype chips we used.

Keywords

  • Chip
  • HPLC
  • Microfluidics
  • Monoliths
  • Peptide separation
  • Protein separation
  • Proteomics
  • Stationary phase

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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