Skip to main content

Protein Purification by Selective Adsorption on Immobilized Pyridinium Ligands

  • Chapter
  • 160 Accesses

Abstract

Avid AL is an affinity gel with synthetic low molecular weight affinity ligands capable of selectively purifying IgG from the serum of all animals tested. By packing the gel in a radial flow column, a high flow rate can be applied and 12 g of highly purified IgG can be obtained within 5 hours directly from the serum. Immunoglobulins bound to Avid AL can be eluted with 0.05 M sodium acetate at pH 3 or at pH 7.4 with a neutral buffer containing “electron-rich” compounds. The affinity gel appears to be suitable under a varieties of storage and depyrogenation conditions.

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

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. R. Lindmark, K. Thoren-Tolling and J. Sjoquist, “Binding of immunoglobulins to Protein A and immunoglobulin levels in mammalian sera.” J. Immunol. Methods, 62, 1–13 (1983).

    Article  PubMed  CAS  Google Scholar 

  2. J. Porath, F. Maisano and M. Belew, “Thiophilic adsorption–a new method for protein fractionation.” FEES Lett., 185, 306–310 (1985).

    Article  CAS  Google Scholar 

  3. M. Belew, N. Juntti, A. Larsson and J. Porath, “A one-step purification method for monoclonal antibodies based on salt-promoted adsorption chromatography on a ”thiophilic“ adsorbent.” J. Immunol. Methods, 102, 173–182 (1987).

    Article  PubMed  CAS  Google Scholar 

  4. T. W. Hutchens and J. Porath, Thiophilic adsorption, “A comparison of model protein behavior.” Biochem., 26, 7199–7204 (1987).

    Article  CAS  Google Scholar 

  5. T. T. Ngo, “Synthetic affinity ligand compositions and methods for purification and recovery of organic molecules.” U.S.Patent 4, 981–961

    Google Scholar 

  6. T. T. Ngo and N. Khatter, “Chemistry and preparation of affinity ligands useful in immunoglobulin isolation and serum protein separation.” J. Chrom., 510, 281–291 (1990).

    Article  CAS  Google Scholar 

  7. T. T. Ngo and N. Khatter, “Rapid and simple isolation of multigram goat IgG from serum using Avid ALTM and radical flow column.” Appl. Biochem. Biotech., 30, 111–119 (1991).

    Article  CAS  Google Scholar 

  8. T. T. Ngo and N. Khatter, “Avid AL, a synthetic ligand affinity gel mimicking immobilized bacterial antibody receptor for purification of immunoglobulin G.” J. Chrom., 597, 101–109 (1992).

    Article  CAS  Google Scholar 

  9. A. R. Katritzky and J. M. Lagowski, The principles of heterocyclic chemistry Methuen & Co. Ltd., London, 1967, pp. 41–76.

    Google Scholar 

  10. T. L. Gilchrist, Heterocyclic Chemistry, Pitman, London, 1985, pp. 247–269.

    Google Scholar 

  11. R. D. Chamber, J. Hutchinson and W. K. R. Musgrave, “Polyfluoroheterocyclic compounds. Part II. Nucleophilic substitution in pentafluopyridine.” J. Chem. Soc., 3736–3739 (1964).

    Google Scholar 

  12. R. E. Banks, J. E. Burgess, W. M. Cheng and R. N. Haszeldine, “Heterocyclic polyfluoro-compounds. Part IV. Nucleophilic substitution in pentafluoropyridine: The preparation and properties of some 4-subsituted 2,3,5,6tetrafluoropyridines.” J. Chem. Soc., 575–581 (1965).

    Google Scholar 

  13. R. D. Chamber and C. R. Sargent, in: Polyfluoroaromatic compounds in: Advances in Heterocyclic Chemistry Volume 28, A. R. Katritzky and A. J. Boulton, Eds., Academic Press, New York, 1982, pp. 47–59.

    Google Scholar 

  14. R. D. Chamber, W. Kenneth, R. Musgrave and P. G. Urben, “Pyridium salts of halogenated heterocyclic compounds.” Chem. Ind., 89 (1975).

    Google Scholar 

  15. J. Porath, “Salting-out adsorption techniques for protein purification.” Biopolymers, 26, 5193–5204 (1987).

    Article  Google Scholar 

  16. G. Cilento and P. Giusti, “Electron transfer from the indole nucleus to the pyridine coenzyme.” J. Am. Chem. Soc., 81, 3801–3802 (1959).

    Article  CAS  Google Scholar 

  17. G. Cilento and P. Tedeschi, “Pyridine coenzymes. IV. Charge transfer interaction with the indole nucleus.” J. Biol. Chem., 236, 907–910 (1961).

    PubMed  CAS  Google Scholar 

  18. S. G. A. Alivisatos, G. A. Mourkides and A. Jibril, “Non-enzymic reactions of indoles with coenzyme I.” Nature, 186, 718–719 (1960).

    Article  PubMed  CAS  Google Scholar 

  19. E. M. Kosower, in: The Enzymes, Volume 3, Charge-transfer complexing of pyridium rings P. D. Boyer, H. Lardy and K. Myrback, Eds., Academic Press, New York, 1960, pp. 171–194.

    Google Scholar 

  20. F. Ungar and S. G. A. Alivisatos, “Spectrophotometric evidence of certain anion-DNP+ interactions (including orthophosphates).” Biochim. Biophys. Acta, 46, 406–408 (1961).

    Article  CAS  Google Scholar 

  21. S. G. A. Alivisatos, F. Ungar, U. A. Jibril and G. A. Mourkides, “Non-enzymic reaction of indole with pyridine coenzymes and related structures.” Biochim. Biophys. Acta, 51, 361–372 (1961).

    Article  PubMed  CAS  Google Scholar 

  22. J. Porath, in: Protein Recognition of Immobilized Ligands T. W. Hutchens, Ed., Alan R. “Electron-donor-acceptor chromatography (EDAC) for biomolecules in aqueous solutions.” Liss Inc., New York, pp. 101–122, 1989.

    Google Scholar 

  23. M. A. Petti, T. J. Shepodd, R. E. Barrans, Jr., and D. A. Dougherty, “Hydrophobic binding of water-soluble guests by high-symmetry, chiral hosts. An electron-rich receptor site with a general affinity for quaternary ammonium compounds and electron-deficient systems.” J. Am. Chem. Soc., 110, 6825–6840 (1988).

    Article  CAS  Google Scholar 

  24. D. A. Dougherty and D. A. Stauffer, “Acetylcholine binding be a synthetic receptor: implications for biological recognition. ” Science, 250, 1558–1560 (1990).

    Article  PubMed  CAS  Google Scholar 

  25. Y. Satow, G. H. Cohen, E. A. Padlan and D. R. Davies, “Phosphocholine binding immunoglobulin Fab McPC603, An X-ray difraction study at 2.7.” J. Mol. Biol., 190, 593–604 (1986).

    Article  PubMed  CAS  Google Scholar 

  26. E. D. Getzoff, J. A. Tainer, R. A. Lerner and H. M. Geysen, in: Advances in Immunology Volume 43, F. J. Dixon, Ed., “The chemistry and mechanism of antibody binding to protein antigens.” Academic Press, San Diego, pp. 1–98, 1988.

    Google Scholar 

  27. J. L. Sussman, M. Harel, F. Frolow, C. Oefner, A. Goldman, L. Toker and I. Silman, “Atomic structure of acetylcholinesterase from Torpedo californica: A prototypic acetylcholine-binding protein.” Science, 253, 872–879 (1991).

    Article  PubMed  CAS  Google Scholar 

  28. D. R. Davies and H. Metzger, “Structural basis of antibody function.” Ann. Rev. Immunol., 1, 87–117 (1983).

    Article  CAS  Google Scholar 

  29. S. K. Burley and Petsko, “Amino-aromatic interactions in proteins.” FEBS Lett., 203, 139–143 (1986).

    Google Scholar 

  30. K. C. S. Reid, P. F. Lindley and J. M. Thornton, “Sulphur-aromatic interactions in proteins. ” FEBS Lett., 190, 209–213 (1985).

    Google Scholar 

  31. J. Porath and M. Belew, “Thiophilic” interaction and the selective adsorption of proteins.“ Trends in Biotech.,” 5, 225–229 (1987).

    Article  CAS  Google Scholar 

  32. J. Porath, “Explorations into the field od charge-transfer adsorption.” J. Chrom., 159, 13–24 (1978).

    Google Scholar 

  33. J. Porath, “Charge-transfer adsorption in aqueous media.” Pure & Applied Chemistry, 51, 1549–1559 (1979).

    Article  CAS  Google Scholar 

  34. D. E. Atkinson, in Current Topics in Cellular Regulation Volume 1, B. L. Horecker and E. R. Stadtman, Eds., “Limitation of metabolite concentrations and the conservation of solvent capacity in the living cell.”, Academic Press, New York, pp. 29–43, 1969.

    Google Scholar 

  35. T. W. Hutchens and J. Porath, “Thiophilic adsorption of immunoglobulins–anaylsis of conditions optimal for selective immobilization and purification.” Anal. Biochem., 159, 217–226 (1986).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Springer Science+Business Media New York

About this chapter

Cite this chapter

Ngo, T.T. (1994). Protein Purification by Selective Adsorption on Immobilized Pyridinium Ligands. In: Gebelein, C.G., Carraher, C.E. (eds) Biotechnology and Bioactive Polymers. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-9519-6_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-4757-9519-6_18

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4757-9521-9

  • Online ISBN: 978-1-4757-9519-6

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics