Skip to main content
SpringerLink
Log in

Separation of biomolecules by μ-high-performance anion-exchange chromatography using a tentacle-like stationary phase

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

Abstract

A variety of biomolecules and their variants, which have previously been problematic to separate, have been analysed using a novel anion-exchange resin based on a non-porous polystyrene polymeric support with a hydrophilic coating and grafted tentacular quaternary ammonium functional groups. The hydrophilic coating results in minimal interaction between the support and the biomolecule, while the highly flexible tentacular-like anion-exchange groups increase the ionic interaction potential and act as an umbrella to hold the proteins away from the surface. Because of the removal of silanophilic interactions, minimisation of hydrophobic interactions, and the highly flexible nature of the tentacle-like ion exchangers, ionic interactions can therefore dominate the separation. As such this phase is highly suited to the separation of highly charged biomolecules and their variants. This polymeric strong anion-exchange (SAX) support was packed into a fused-silica capillary column and, using a salt concentration gradient, various recombinant proteins were analysed by μ-HPLC resulting in baseline resolution.

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

Abbreviations

MW:

Molecular weight

MS:

Mass spectrometry

HPLC:

High-performance liquid chromatography

SAX:

Strong anion exchange

rV Antigen:

Recombinant V antigen

rNAPc2:

Recombinant Nenatode Anticoagulant Protein C2

kDa:

Kilo Daltons

pI:

Isoelectric point

TF:

Tissue factor

FVlla:

Factor Vlla

Tris:

Tris(hydroxymethyl)methylamine

References

  1. Weitzhandler M, Farnan D, Rohrer JS, Avdalovic N (2001) Proteomics 1:179–185

    Article  CAS  PubMed  Google Scholar 

  2. Weitzhandler M, Farnan D, Horvath J, Rohrer JS, Slingsby RW, Avdalovic N, Pohl C (1998) J Chromatogr A 365–372

  3. Zhang J, Huang X, Zhang S, Horvath C (2000) Anal Chem 72:3022–3029

    Article  CAS  PubMed  Google Scholar 

  4. Legido-Quigley C, Smith NW, Mallet D (2002) J Chromatogr A 976:11–18

    Article  CAS  PubMed  Google Scholar 

  5. Premstaller A, Oberacher H, Huber CG (2000) Anal Chem 72:4386–4393

    Article  CAS  PubMed  Google Scholar 

  6. Premstaller A, Oberacher H, Walcher W, Timperio AM, Zolla L, Chervet JP, Cavusoglu N, van Dorsselaer A, Huber CG (2001) Anal Chem 73:2390–2396

    Article  CAS  PubMed  Google Scholar 

  7. Muller W (1990) J Chromatogr A 133–140

  8. Zou H, Huang X, Ye M, Luo Q (2002) J Chromatogr A (Review of mono’s) 954:5–32

    Article  CAS  Google Scholar 

  9. Legido-Quigley C, Marlin ND, Melin V, Manz A, Smith NW (2003) Electrophoresis 24:917–944

    Article  CAS  PubMed  Google Scholar 

  10. Legido-Quigley C, Marlin ND, Smith NW (2004) J Chromatogr A 1030:195–200

    Article  CAS  PubMed  Google Scholar 

  11. Horvath CS, Lipsky S (1969) J Chromatogr Sci 7:109–116

    CAS  Google Scholar 

  12. Lee WC (1997) J Chromatogr B 699:29–45

    CAS  Google Scholar 

Download references

Acknowledgements

The authors thank Avecia Ltd (Blackley, UK) for funding the project and providing samples.

Author information

Authors and Affiliations

  1. Centre for Analytical Sciences, Department of Chemistry, Imperial College London, Exhibition Road, London, SW7 2AY, UK

    Nicola Dawn Marlin

  2. Microseparations Group, Department of Pharmacy, King’s College London, Franklin-Wilkins Building, 150 Stamford Street, London, SE1 9NH, UK

    Norman William Smith

Authors
  1. Nicola Dawn Marlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Norman William Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nicola Dawn Marlin.

Additional information

Awarded a poster prize on the occasion of the Euranalysis XIII conference, Salamanca, Spain, 5–10 September 2004

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marlin, N.D., Smith, N.W. Separation of biomolecules by μ-high-performance anion-exchange chromatography using a tentacle-like stationary phase. Anal Bioanal Chem 382, 493–497 (2005). https://doi.org/10.1007/s00216-004-2944-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-004-2944-z

Keywords

Search

Navigation