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Biochemistry (Moscow)

, Volume 74, Issue 2, pp 162–164 | Cite as

Adsorption chromatography of proteins

  • V. I. Surovtsev
  • V. M. BorzenkovEmail author
  • K. V. Detushev
Article

Abstract

A method for adsorption chromatography of proteins is proposed. A protein solution is passed through a cellulose column at a pH value corresponding to an isoelectric point of the protein. Depending on the charge of unwanted proteins, they either remain at the origin (if charges of protein and ion-exchanger are opposite) or are released from the column (if charges of protein and ion-exchanger coincide). Elution volume of the purified protein is higher than for the second group of unwanted proteins because movement of the uncharged protein of interest includes its adsorption on cellulose followed by subsequent desorption caused by the elution buffer. Problems of optimization of buffers and adsorbents are discussed. Applicability of the method of adsorption chromatography is illustrated using purification of horseradish peroxidase as an example.

Key words

adsorption chromatography adsorbent-ion exchanger cellulose buffer peroxidase amine 

Abbreviations

MOPS

3-(N-morpholino)propanesulfonic acid

MOPSO

3-(N-morpholino)-2-hydroxypropanesulfonic acid

TEA

triethanolamine

TES

N-tris(hydroxymethyl)methyl-2-aminosulfonic acid

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References

  1. 1.
    Tsvet, M. S. (1946) Chromatographic Adsorption Analysis [in Russian], USSR Academic Press, Moscow, pp. 9–39.Google Scholar
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    Osterman, L. A. (1985) Chromatography of Proteins and Nucleic Acid [in Russian], Nauka, Moscow, pp. 221–272.Google Scholar
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    Dolman, D., Newell, G. A., Thurlow, M. D., and Dunford, H. B. (1975) Can. J. Biochem., 53, 495–500.PubMedCrossRefGoogle Scholar
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    Berezin, I. V., Ugarova, N. N., Kershengol’tz, B. M., and Brovko, L. Yu. (1975) Biokhimiya, 40, 297–301.Google Scholar
  5. 5.
    Scopes, R. (1985) Methods of Protein Purification [Russian translation], Mir, Moscow, pp. 243–247.Google Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • V. I. Surovtsev
    • 1
  • V. M. Borzenkov
    • 1
    Email author
  • K. V. Detushev
    • 1
  1. 1.State Research Center of Applied Microbiology and BiotechnologyMinistry of Health and Social Development of the Russian FederationObolensk, Moscow RegionRussia

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