Analytical and Bioanalytical Chemistry

, Volume 397, Issue 8, pp 3421–3430

New ammunition for the proteomic reactor: strong anion exchange beads and multiple enzymes enhance protein identification and sequence coverage

  • Hu Zhou
  • Weimin Hou
  • Jean-Philippe Lambert
  • Daniel Figeys
Original Paper

DOI: 10.1007/s00216-010-3791-8

Cite this article as:
Zhou, H., Hou, W., Lambert, JP. et al. Anal Bioanal Chem (2010) 397: 3421. doi:10.1007/s00216-010-3791-8
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Abstract

The enrichment and processing of proteomic samples prior to multi-dimensional chromatography remain a challenge in ‘gel-free’ proteomics. We previously reported the development of a microfluidic device called the “proteomic reactor” that relied on enriching proteins by using strong cation exchange (SCX) followed by trypsin digestion in an interstitial volume as little as 50 nL. Here, we report a novel proteomic reactor that is based on polymeric strong anion exchange (SAX) material to analyse proteomic samples. We also compare the performance of the SAX proteomic reactor to our previously reported SCX proteomic reactor for analysing complex yeast proteomes. Our results indicate that the SAX protein reactor preferentially identifies more acidic peptides and proteins compared to the SCX reactor. We show that the SAX and SCX reactors are complementary and that their combination increases the number of unique peptides and proteins identified by 50%. Furthermore, we show that the number of protein identified can be increased further by up to 40% using different proteolytic enzymes on the proteomic reactor.

Keywords

Proteomic reactor Strong cation exchange Strong anion exchange Protein coverage Microfluidic 

Supplementary material

216_2010_3791_MOESM1_ESM.pdf (1.1 mb)
ESM 1(PDF 1.05 MB)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Hu Zhou
    • 1
    • 2
  • Weimin Hou
    • 1
    • 3
  • Jean-Philippe Lambert
    • 1
    • 2
  • Daniel Figeys
    • 1
    • 2
    • 3
  1. 1.Ottawa Institute of Systems BiologyUniversity of OttawaOttawaCanada
  2. 2.Department of Biochemistry, Microbiology and Immunology, Faculty of MedicineUniversity of OttawaOttawaCanada
  3. 3.Department of Chemistry, Faculty of ScienceUniversity of OttawaOttawaCanada

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