Affinity Chromatography — Old Problems and New Approaches

  • Steven C. March
  • Indu Parikh
  • Pedro Cuatrecasas
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 42)


The purification of macromolecules by biospecific adsorption-affinity chromatography (1) is a technique with a long history although its application to a wide variety of problems is comparatively recent. As long ago as 1910 there were reports of the purification of amylase by adsorption to insoluble starch (2). Tyrosinase was purified in 1953 using a cellulose matrix bearing the inhibitor, diazodizine (3). In both of these examples the power of purification lies in the highly selective “insolubilization” of the macromolecule from solution. This reversible phase separation often allows a high degree of concentration of the particular molecular species of interest from dilute solution and the removal, by washing and elution processes, of other molecular species that are cosolutes.


Affinity Chromatography Cyanogen Bromide Reversible Phase Separation Aleutian Mink Disease Virus Affinity Chroma 
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  1. 1.
    Cuatrecasas, P. and Anfinsen, C. B. “Affinity Chromatography” An. Rev. Biochem. 40: 259, 1971.CrossRefGoogle Scholar
  2. 2.
    Starkenstein, E. “Uber Fermentwirkung and deren Besiuflussung durch Neutralsalze” Biochem. Z. 24: 210, 1910.Google Scholar
  3. 3.
    Lerman, L. S. “A Biochemically Specific Method for Enzyme Isolation” Proc. Natl. Acad. Sci. USA 39: 232, 1953.CrossRefGoogle Scholar
  4. 4.
    Ashani, Y. and Wilson, I. B. “A Covalent Affinity Column for the Purification of Acetylcholine-esterase” Biochim. Biophys. Acta 276: 317, 1972.Google Scholar
  5. 5.
    Axen, R. and Parath, J. “Chemical Coupling of Enzymes to Cross-Linked Dextran (Sephadex)” Nature 210: 367, 1966.CrossRefGoogle Scholar
  6. 6.
    Cuatrecasas, P., Wilchek, M., Anfinsen, C. B. “Selective Enzyme Purification by Affinity Chromatography” Proc. Natl. Acad. Sci. USA 61: 636, 1968.CrossRefGoogle Scholar
  7. 7.
    Steers, E., Cuatrecasas, P., and Pollard, H. “The Purification of (3-Galactosidase from Escherichia coli by Affinity Chromatography” J. Biol. Chem. 246: 196, 1971.Google Scholar
  8. 8.
    Sica, V., Nola, E., Parikh, I., Puca, G. A., and Cuatrecasas, P. “Purification of Oestradiol Receptors by Affinity Chromatography” Nature New Biology 244: 36, 1973.Google Scholar
  9. 9.
    Cuatrecasas, P. “Interaction of Insulin with Cell Membrane-Primary Action of Insulin” Proc. Natl. Acad. Sci. USA 63: 450, 1969.CrossRefGoogle Scholar
  10. 10.
    Soderman, D. D., Germershausen, J. and Katzen, H. M. “Affinity Binding of Intact Fat Cells and their Ghosts to Immobilized Insulin” Proc. Natl. Acad. Sci. USA 70: 792, 1973.CrossRefGoogle Scholar
  11. 11.
    Henry, C., Kimura, J., and Wofsy, L. “Cell Separations on Affinity Columns - Isolation of Immunospecific Precursor Cells from Unimmunized Mice” Proc. Natl. Acad. Sci. USA 69: 34, 1972.CrossRefGoogle Scholar
  12. 12.
    Truffa-Bachi, P. and Wofsy, L. “Specific Separation of Cells by Affinity Chromatography” Proc. Natl. Acad. Sci. USA 66: 685, 1970.CrossRefGoogle Scholar
  13. 13.
    Singhal, S. K. and Wigzell, H. “In Vitro Induction of Specific Unresponsiveness of Immunologically Reactive Normal Bone Marrow Cells” J. Exptl. Med. 131: 149, 1970.CrossRefGoogle Scholar
  14. 14.
    Davie, J. M. and Paul, W. F. “Receptors on Immunocompetent Cells. I. Receptor Specificity of Cells Participating in a Cellular Immune Response” Cell. Immunol. 1: 404, 1970.CrossRefGoogle Scholar
  15. 15.
    Rutishauer, U., Millette, C. F. and Edelman, G. M. “Specific Fractionation of Immune Cell Populations” Proc. Natl. Acad. Sci. USA 69: 1596, 1972.CrossRefGoogle Scholar
  16. 16.
    Miller, J. V., Cuatrecasas, P., and Thompson, E. B. “Partial Purification by Affinity Chromatography of Tyrosine Aminotransferase-Synthesizing Ribosomes from Hepatoma Tissue Culture Cells” Proc. Natl. Acad. Sci. USA 68: 1014, 1971.CrossRefGoogle Scholar
  17. 17.
    Kenyon, A. J., Gander, J. E., Lopez, C., and Good, R. A. “Isolation of Aleutian Mink Disease Virus by Affinity Chromatography” Science 179: 187, 1973.CrossRefGoogle Scholar
  18. 18.
    Cuatrecasas, P. and Illiano, G. “Purification of Neuraminidase from Vibrio Cholerea, Clostridium and Influenza Virus by Affinity Chromatography” Biochem. Biophys. Res. Comm. 44: 178, 1971.CrossRefGoogle Scholar
  19. 19.
    Cuatrecasas, P. and Tell, G. P. E. “Insulin-Like Activity of Concanavalin A and Wheat-Germ Agglutinin - Direct Interactions with Insulin Receptors” Proc. Natl. Acad. Sci. USA 70: 485, 1973.CrossRefGoogle Scholar
  20. 20.
    Steineman, A. and Stryer, L. “Accessibility of the Carbohydrate Moiety of Rhodopsin” Biochemistry 12: 1499, 1973.CrossRefGoogle Scholar
  21. 21.
    Cuatrecasas, P. “Isolation of Insulin Receptor of Liver and Fat-Cell Membranes” Proc. Natl. Acad. Sci. USA 69: 316, 1972.Google Scholar
  22. 22.
    Sica, V., Parikh, I., Nola, F., Puca, G. A., and Cuatrecasas, P. “Affinity Chromatography and the Purification of Estrogen Receptors” J. Biol. Chem. 248: 6543, 1973.Google Scholar
  23. 23.
    Allen, R. H. and Majerus, P. W. “Isolation of Vitamin B12-Binding Proteins Using Affinity Chromatography” J. Biol. Chem. 247: 7695, 1972.Google Scholar
  24. 24.
    Venter, J. C., Ross, J., Dixon, J. E., Mayer, S. E. and Kaplan, N. O. “Immobilized Catecholamines and Cocaine Effects on Contractility of Cardiac Muscle” Proc. Natl. Acad. Sci. USA 70: 1214, 1973.CrossRefGoogle Scholar
  25. 25.
    Erel, Z., Zaidenzaig, Y., and Shaltiel, S. “Hydrocarbon Coated Sepharoses. Use in the Purification of Glycogen Phosphorylase” Biochem. Biophys. Res. Comm. 49: 383, 1972.CrossRefGoogle Scholar
  26. 26.
    Axen, R., Porath, M., and Ernback, S. “Chemical Coupling of Peptides and Proteins to Polysaccharides by Means of Cycrogen Halids” Nature 214: 1302, 1967.CrossRefGoogle Scholar
  27. 27.
    Svensson, B. “Use of Isoelectric Focusing to Characterize the Bonds Established During Coupling of CNBr-Activated Amylodextrin to Subtilisin Type Novo” FEBS Letters 29: 167, 1973.CrossRefGoogle Scholar
  28. 28.
    Cuatrecasas, P. Protein Purification by Affinity Chromatography. Derivatizations of Agarose and Polyacrylamide Beads. J. Biol. Chem. 245: 3059, 1970.Google Scholar
  29. 29.
    Inman, J. K. and Dintzis, H. The Derivatization of Cross-Linked Polyacrylamide Beads. Controlled Introduction of Functional Groups for the Preparation of Special-Purpose, Biochemical Adsorbents. Biochemistry 10: 4074, 1969.CrossRefGoogle Scholar
  30. 30.
    Parikh, I., March, S. and Cuatrecasas, P. “Advances in the Methodology of Substitution Reactions and in the Use of Agarose in Affinity Chromatography” Methods in Enzymology 22 (B) Affinity Techniques (ed. W. Jakoby and M. Wilchek) Academic Press (in press).Google Scholar
  31. 31.
    Oka, T., Topper, Y. J. “Dynamics of Insulin Action on Mammary Epithelium” Nature New Biology 239: 216, 1972.Google Scholar
  32. 32.
    Lowe, C. R., Harvey, M. J., Craven, D. B., Kerfoot, M. A., Hollows, M. F., and Dean, P. D. G. “The Purification of Nicotinamide Nucleotide-Dependent Dehydrogenases on Immobilized Cofactors” Biochem. J. 133: 507, 1973.Google Scholar
  33. 33.
    Cuatrecasas, P. and Parikh, I. “Adsorbents for Affinity Chromatography. Use of N-Hydroxysuccinimide Esters of Agarose” Biochem. 11: 229, 1972.CrossRefGoogle Scholar
  34. 34.
    BIO-RAD Laboratories, 32nd and Griffin Ave., Richmond, Ca. 94804.Google Scholar
  35. 35.
    Pharmacia Fine Chemicals Inc., 800 Centennial Ave., Piscataway, N. J. 08854.Google Scholar
  36. 36.
    March, S. C., Parikh, I., and Cuatrecasas, P. “A Simplified Method for Cyanogen Bromide Activation of Agarose for Affinity Chromatography” An. Biochem. (in press).Google Scholar

Copyright information

© Plenum Press, New York 1974

Authors and Affiliations

  • Steven C. March
    • 1
  • Indu Parikh
    • 1
  • Pedro Cuatrecasas
    • 1
  1. 1.Johns Hopkins University School of MedicineBaltimoreUSA

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