Hydrophobic Interaction Chromatography of Proteins on Neutral Adsorbents

  • Stellan Hjertén
Part of the Biological Separations book series (BIOSEP)


The separation mechanisms of most of the high-resolution biochemical fractionation techniques are based mainly on one particular separation parameter: in nonsieving electrophoresis and ion exchange chromatography, the surface charge of the solute of interest; in ultracentrifugation, in chromatographic molecular sieving, and in pore-gradient electrophoresis, the molecular size; in isopycnic gradient centrifugation, the density; in hydroxylapatite chromatography, the number of available carboxylic (and phosphate) groups in the solute molecules (Bernardi and Kawasaki, 1968). Since the above methods utilize mainly one separation parameter, they often give reproducible results, and the separation patterns can in many cases be correlated to particular molecular properties of the fractionated substances. For this reason, these separation methods are used routinely in most biochemical research laboratories. However, two or more separation parameteres can with advantage be utilized simultaneously, provided that the contribution of each parameter can be kept constant from experiment to experiment, which is a prerequisite for reproducible results. In practice, it is often very difficult to fulfill this requirement. Polyacrylamide gel electrophoresis is an example of the few methods which utilize two separation parameters and still provide both high reproducibility and high resolution.


Glycidyl Ether Hydrophobic Interaction Chromatography Cyanogen Bromide Separation Parameter Separation Pattern 
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  1. Axén, R., Porath, J., and Ernback, S. (1967). Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature (London) 214:1302.CrossRefGoogle Scholar
  2. Bernardi, G., and Kawasaki, T. (1968). Chromatography of polypeptides and proteins on hydroxyapatite columns. Biochim. Biophys. Acta 160:301.PubMedCrossRefGoogle Scholar
  3. Edelstein, S. J. (1973). Introductory Biochemistry, Holden-Day, San Francisco, p. 130.Google Scholar
  4. Ellingboe, J., Nyström, E., and Sjövall, J. (1970). Liquid-gel chromatography on lipophilic-hydrophobic Sephadex derivatives. J. Lipid Res. 11:266.PubMedGoogle Scholar
  5. Hammar, L., Påhlman, S., and Hjertén S. (1975). Chromatographic purification of a mammalian histidine decarboxylase on charged and noncharged alkyl derivatives of agarose. Biochim. Biophys. Acta 403:554.PubMedCrossRefGoogle Scholar
  6. Hjertén, S. (1973). Some general aspects of hydrophobic interaction chromatography. J. Chromatogr. 87:325.CrossRefGoogle Scholar
  7. Hjertén, S. (1976). Purification of proteins by hydrophobic interaction chromatography. In Protides of the Biological Fluids, Vol. 23 (H. Peeters, ed.), Pergamon, New York (in press).Google Scholar
  8. Hjertén, S., and Johansson, K.-E. (1972). Selective solubilization with Tween 20 of membrane proteins from Acholeplasma laidlawii. Biochim. Biophys. Acta 288:312.CrossRefGoogle Scholar
  9. Hjertén, S., Rosengren, J., and Påhlman, S. (1974). Hydrophobic interaction chromatography The synthesis and the use of some alkyl and aryl derivatives. J. Chromatogr. 101:281.CrossRefGoogle Scholar
  10. Hofstee, B. H. J. (1973a). Immobilization of enzymes through noncovalent binding to substituted agaroses. Biochem. Biophys. Res. Commun. 53:1137.PubMedCrossRefGoogle Scholar
  11. Hofstee, B. H. J. (1973b). Hydrophobic affinity chromatography of proteins. Anal. Biochem. 52:430.PubMedCrossRefGoogle Scholar
  12. Lewin, S. (1974). Displacement of Water and Its Control of Biochemical Reactions, Academic Press, New York.Google Scholar
  13. Rosengren, J., Påhlman, S., Glad, M., and Hjertén, S. (1975). Hydrophobic interaction chromatography on noncharged Sepharose derivatives: Binding of a model protein related to ionic strength, hydrophobicity of the substituent, and degree of substitution (determined by NMR). Biochim. Biophys. Acta 412:51.PubMedCrossRefGoogle Scholar
  14. Tanford, C. (1973). The Hydrophobic Effect: Formation of Micelles and Biological Membranes, Wiley, New York.Google Scholar
  15. Ulbrich, V., Makeš, J., and Jureček, M. (1964). Identifizierung der Glycidyläther Bis-Phenyl- und Bis-α-Naphthylurethane der α-Alkyl(Aryl)äther des Glycerins. Collect. Czech. Chem. Commun. 29:1466.Google Scholar

Copyright information

© Springer Science+Business Media New York 1976

Authors and Affiliations

  • Stellan Hjertén
    • 1
  1. 1.Institute of Biochemistry, Biomedical CenterUniversity of UppsalaUppsalaSweden

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