Skip to main content
SpringerLink
Log in

Investigation of sample-purification procedures for MALDI-based proteomic studies

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Purification methods for proteomics samples are of crucial concern for improving the quality of the sample delivered to the mass spectrometer. They constitute the link between the mass spectrometer and protein processing and peptide isolation steps that usually require solvents, buffers, or detergents completely incompatible with MS-analysis conditions. This work describes three new clean-up procedures using synthetic membranes and polymer media and compares them with standard procedures. The efficiency of each of the purification procedures was studied via application to four standards and two membrane proteins. This work highlights the importance of versatility in sample preparation, especially for MS-based proteomic investigations.

PMF spectra obtained after MALDI-TOF measurements of bovine mitochondrial complex III (A) and complex IV (B) in-solution digests, with and without purification

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Patterson SD, Aebersold RH (2003) Nature Genetics Suppl 33:311–323

    Article  CAS  Google Scholar 

  2. Karas M, Bachmann D, Bahr U, Hillenkamp F (1987) Int J Mass Spectrom Ion Proc 78:53–68

    Article  CAS  Google Scholar 

  3. Karas M, Hillenkamp F (1988) Anal Chem 60:2299–2301

    Article  CAS  Google Scholar 

  4. Xu Y, Bruening ML, Watson JT (2003) Mass Spectrom Rev 22:429–440

    Article  CAS  Google Scholar 

  5. Ishihama Y (2005) J Chromatogr A 1067:73–83

    Article  CAS  Google Scholar 

  6. Chen G, Pramanik BN, Liu YH, Mirza UA (2007) J Mass Spectrom 42:279–287

    Article  CAS  Google Scholar 

  7. Le Gac S, Carlier J, Camart JC, Cren-Olivé C, Rolando CJ (2004) J Chromatogr B 808:3–14

    Article  Google Scholar 

  8. Rappsilber J, Ishihama Y, Mann M (2003) Anal Chem 75:663–670

    Article  CAS  Google Scholar 

  9. Kussman M, Nordhoff E, Rahbek-Nielsen H, Haebel S, Rossel-Larsen M, Jakobsen L, Gobom J, Mirgorodskaya E, Kroll-Kristensen A, Palm L, Roepstorff P (1997) J Mass Spectrom 32:593–601

    Article  Google Scholar 

  10. Gobom J, Nordhoff E, Ekman R, Roepstorff P (1997) Int J Mass Spectrom Ion Proc 169/170:153–163

    Article  CAS  Google Scholar 

  11. Gobom J, Nordhoff E, Mirgorodskaya E, Ekman R, Roepstorff P (1999) J Mass Spectrom 34:105–116

    Article  CAS  Google Scholar 

  12. Havlis J, Thomas H, Sebela M, Shevchenko A (2003) Anal Chem 75:1300–1306

    Article  CAS  Google Scholar 

  13. Fandiño A, Rais I, Vollmer M, Elgass H, Schägger H, Karas M (2005) J Mass Spectrom 40:1223–1231

    Article  Google Scholar 

  14. Winston RL, Fitzgerald MC (1998) Anal Biochem 262:83–85

    Article  CAS  Google Scholar 

  15. Worrall TA, Cotter RJ, Woods AS (1998) Anal Chem 70:750–756

    Article  CAS  Google Scholar 

  16. Bull HB, Breese K (1974) Arch Bioch Biophys 161:665–670

    Article  CAS  Google Scholar 

  17. Kyte J, Doolittle RF (1988) J Mol Biol 157:105–132

    Article  Google Scholar 

  18. Tannu NS, Wu J, Rao VK, Gadgil HS, Pabst MJ, Gerling IC, Raghow R (2004) Anal Biochem 327:222–232

    Article  CAS  Google Scholar 

  19. Zhang N, Li L (2004) Rapid Commun Mass Spectrom 18:889–896

    Article  CAS  Google Scholar 

  20. Schägger H, Pfeiffer K (2000) EMBO J 19:1777–1783

    Article  Google Scholar 

Download references

Acknowledgements

We thank I. Rais and H. Schägger (Zentrum der Biologischen Chemie, Universitätsklinikum, Frankfurt am main, Germany) who provided the blue native gel bands of membrane protein complexes.

Author information

Authors and Affiliations

  1. Institut für Pharmazeutische Chemie, Johann Wolfgang Goethe Universität, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Marianne André & Michael Karas

Authors
  1. Marianne André
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael Karas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael Karas.

Rights and permissions

Reprints and permissions

About this article

Cite this article

André, M., Karas, M. Investigation of sample-purification procedures for MALDI-based proteomic studies. Anal Bioanal Chem 389, 1047–1053 (2007). https://doi.org/10.1007/s00216-007-1471-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-007-1471-0

Keywords

Search

Navigation