Skip to main content
SpringerLink
Log in

Gelatin-Immobilised Poly(hydroxyethyl methacrylate) Cryogel for Affinity Purification of Fibronectin

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

In this work, fibronectin purification from human plasma with the gelatin-immobilised poly(hydroxyethyl methacrylate) (PHEMA) cryogel has been evaluated. The PHEMA cryogel was prepared by cryo-polymerisation which proceeds in an aqueous solution of monomer frozen inside a plastic syringe. The PHEMA cryogel contained interconnected macrochannels of 10–200 μm in diameter. Gelatin molecules were covalently immobilised onto the PHEMA cryogel via carbodiimide activation. The gelatin-immobilised PHEMA cryogel was used to purify fibronectin from human plasma. Fibronectin adsorption from human plasma on the PHEMA cryogel was 0.30 mg/ml, while much higher adsorption values, up to 38 mg/ml, was obtained with the gelatin-immobilised PHEMA cryogel. The fibronectin adsorption capacity of the gelatin-immobilised PHEMA cryogel did not change with an increase in the flow rate of plasma. Up to 92 % of the adsorbed fibronectin was eluted using 2 M urea containing 1 M NaCl as elution agent. The adsorption–elution cycle was repeated ten times using the same PHEMA cryogel. No remarkable decrease was detected in the adsorption capacity of the gelatin-immobilised PHEMA cryogel.

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
Fig. 6

Similar content being viewed by others

References

  1. Hynes, R. O. (1990). Fibronectin. New York: Springer.

    Book  Google Scholar 

  2. Ruoslahti, E., & Reed, J. C. (1994). Cell, 77, 477–478.

    Article  CAS  Google Scholar 

  3. Regnault, V., Rivat, C., & Stoltz, J. F. (1988). Journal of Chromatography, 432, 93–102.

    CAS  Google Scholar 

  4. Mosesson, M. W., & Umfleet, R. A. (1970). The Journal of Biological Chemistry, 245, 5728–5736.

    CAS  Google Scholar 

  5. Allain, J. P., Lejars, A., Pham, C. T., Gaillandre, A., & Lee, H. H. (1983). Revue Française de Transfusion et Immuno-Hématologie, 26, 123–134.

    Article  CAS  Google Scholar 

  6. Speziale, P., Visai, L., Rindi, S., & Di Poto, A. (2008). Nature Protocol., 3, 525–533.

    CAS  Google Scholar 

  7. Chifflet, S., Bolatto, C., & Tolosa, S. (2004). Journal of Biochemical and Biophysical Methods, 59, 139–143.

    Article  CAS  Google Scholar 

  8. Stol, M., Adam, M., & Deyl, Z. (1987). Journal of Chromatography, 419, 308–314.

    CAS  Google Scholar 

  9. Habeeb, A. F. S. A. (1981). BBA Gen Subjects, 673, 527–538.

    Article  CAS  Google Scholar 

  10. Ruoslahti, E., & Engvall, E. (1980). BBA Gen Subjects, 631, 350–358.

    Article  CAS  Google Scholar 

  11. Ruoslahti, E., Hayman, E. G., Pierschbacher, M., & Engvall, E. (1982). Methods in Enzymology, 82, 803–831.

    Article  CAS  Google Scholar 

  12. Babaç, C., Yavuz, H., Galaev, I. Y., Pişkin, E., & Denizli, A. (2006). React. Functl. Polym., 66, 1263–1271.

    Article  Google Scholar 

  13. Plieva, F. M., Ekström, P., Galaev, I. Y., & Mattiasson, B. (2008). Soft Matter, 4, 2418–2428.

    Article  CAS  Google Scholar 

  14. Alkan, H., Bereli, N., Baysal, Z., & Denizli, A. (2009). Biochemical Engineering Journal, 45, 201–208.

    Article  CAS  Google Scholar 

  15. Arvidsson, P., Plieva, F. M., Savina, I. N., Lozinsky, V. I., Fexby, S., Bülow, L., et al. (2002). Journal of Chromatography. A, 977, 27–38.

    Article  CAS  Google Scholar 

  16. Henderson, T. M. A., Ladewig, K., Haylock, D. N., Mclean, K. M., & O’Connor, A. J. (2013). J. Mater. Chem. B., 1, 2682–2695.

    Article  CAS  Google Scholar 

  17. Gun’ko, V. M., Savina, I. N., & Mikhalovsky, S. V. (2013). Advances in Colloid and Interface Science, 187–188, 1–46.

    Article  Google Scholar 

  18. Reichelt, S., Abe, C., Hainich, S., Knolle, W., Decker, U., Prager, A., et al. (2013). Soft Matter, 9, 2484–2492.

    Article  CAS  Google Scholar 

  19. Sun, S., Tang, Y., Fu, Q., Liu, X., Guo, L., Zhao, Y., et al. (2012). International Journal of Biological Macromolecules, 50, 1002–1007.

    Article  CAS  Google Scholar 

  20. Bereli, N., Şener, G., Altıntaş, E. B., Yavuz, H., & Denizli, A. (2010). Materials Science and Engineering: C, 30, 323–329.

    Article  CAS  Google Scholar 

  21. Perçin, I., Sağlar, E., Yavuz, H., Aksöz, E., & Denizli, A. (2011). International Journal of Biological Macromolecules, 48, 577–582.

    Article  Google Scholar 

  22. Özgür, E., Bereli, N., Türkmen, D., Ünal, S., & Denizli, A. (2011). Materials Science and Engineering: C, 31, 915–920.

    Article  Google Scholar 

  23. Bereli, N., Şener, G., Yavuz, H., & Denizli, A. (2011). Materials Science and Engineering: C, 31, 1078–1083.

    Article  CAS  Google Scholar 

  24. Çimen, D., & Denizli, A. (2012). Colloids Surf B: Biointerfaces, 93, 29–35.

    Article  Google Scholar 

  25. Andaç, M., Galaev, I. Y., & Denizli, A. (2012). Journal of Separation Science, 35, 1173–1182.

    Article  Google Scholar 

  26. Denizli, A. (2002). Journal of Chromatography B, 772, 357–367.

    Article  CAS  Google Scholar 

  27. Harlow, E., & Lane, D. (1988). Antibodies: a laboratory manual. New York: Cold Spring Harbor Laboratory.

    Google Scholar 

  28. Scher, B. M., Mistry, S. J., Haque, N. S., & Scher, W. (2010). Experimental Cell Research, 316, 2402–2413.

    Article  CAS  Google Scholar 

  29. Kashiwagi, S., Nakamura, K., Takeo, K., Takasago, T., Uchimichi, A., & Ito, H. (1991). Electrophoresis, 12, 420–424.

    Article  CAS  Google Scholar 

  30. Vlierberghe, S. V., Vanderleyden, E., Dubruel, P., Vos, F. D., & Schacht, E. (2009). Macromolecular Bioscience, 9, 1105–1115.

    Article  Google Scholar 

  31. Le Van, M. D., & Vermeulen, T. (1981). Journal of Physical Chemistry, 85, 3247–3250.

    Article  Google Scholar 

  32. Cheung, C. W., Porter, J. F., & McKay, G. (2001). Water Research, 35, 605–612.

    Article  CAS  Google Scholar 

  33. Gmeiner, B., Leibl, H., Zerlauth, G., & Seelos, C. (1995). Archives of Biochemistry and Biophysics, 321, 40–42.

    Article  CAS  Google Scholar 

  34. Müller-Schulte, D., Manjini, S., & Vijayalakshmi, M. A. (1991). Journal of Chromatography. A, 539, 307–314.

    Article  Google Scholar 

  35. Smilenov, L., Forsberg, E., Zeligman, I., Sparrman, M., & Johansson, S. (1992). FEBS., 302, 227–230.

    Article  CAS  Google Scholar 

  36. Guo, W., & Ruckenstein, E. (2003). Journal of Chromatography B, 795, 61–72.

    Article  CAS  Google Scholar 

  37. Ho, C. H., Hlady, V., Nyquist, G., Andrade, J. D., & Caldwell, K. D. (1991). Journal of Biomedical Materials Research, 25, 423–441.

    Article  CAS  Google Scholar 

  38. Denizli, F. K., Arıca, M. Y., & Denizli, A. (2001). Journal of Biomaterials Science. Polymer Edition, 12, 479–489.

    Article  Google Scholar 

  39. Poulouin, L., Gallet, O., Rouahi, M., & Imhoff, J. M. (1999). Protein Express Purif, 17, 146–152.

    Article  CAS  Google Scholar 

  40. Alvarez, G. R., Cabrera, I. M., Rodriguez, A. G., Maura, R. B., Torres, A. M., Collazo, M. A. P., et al. (2007). Latin Am J Pharm, 26, 165–170.

    Google Scholar 

  41. Arginine-Sepharose 4B Affinity Chromatography Booklet, Pharmacia Fine Chemicals, 1982.

  42. Gelatin-Sepharose 4B Affinity Chromatography Booklet, Pharmacia Fine Chemicals, 1982.

  43. Matejtschuk, P. (Ed.) (1997). In: Affinity separations: A practical approach (pp. 62–67). Oxford: Oxford University Press.

  44. Retta, S. F., Ferraris, P., & Tarone, G. (1999). In E. Dejans & M. Corada (Eds.), Adhesion protein protocols, vol. 96: Methods in molecular biology (pp. 119–124). Totowa: Humana.

    Chapter  Google Scholar 

  45. Kale, S., & Lali, A. (2011). Biotechnology Progress, 27, 1078–1090.

    Article  CAS  Google Scholar 

  46. Dainiak, M. B., Plieva, F. M., Galaev, I. Y., Kaul, R. H., & Mattiasson, B. (2005). Biotechnology Progress, 21, 644–649.

    Article  CAS  Google Scholar 

  47. Sun, S., Tang, Y., Fu, Q., Liu, X., Du, W., Guo, L., et al. (2012). Journal of Separation Science, 35, 893–900.

    Article  Google Scholar 

  48. Plieva, F. M., Kirsebom, H., & Mattiasson, B. (2011). Journal of Separation Science, 34, 2164–2172.

    CAS  Google Scholar 

  49. Bereli, N., Andaç, M., Baydemir, G., Say, R., Galaev, I. Y., & Denizli, A. (2008). Journal of Chromatography. A, 1190, 18–26.

    Article  CAS  Google Scholar 

  50. Bereli, N., Ertürk, G., & Denizli, A. (2012). Separation Science and Technology, 47, 1813–1820.

    Article  CAS  Google Scholar 

  51. Plieva, F. M., Galaev, I. Y., & Mattiasson, B. (2007). Journal of Separation Science, 30, 1657–1671.

    Article  CAS  Google Scholar 

  52. Perçin, I., Yavuz, H., Aksöz, E., & Denizli, A. (2012). Biotechnology Progress, 28, 756–761.

    Article  Google Scholar 

Download references

Acknowledgement

This study was supported by Scientific Research and Development Office of Hacettepe University (010D12601003).

Author information

Authors and Affiliations

  1. Department of Biology, Molecular Biology Division, Hacettepe University, Beytepe, 06800, Ankara, Turkey

    Işık Perçin & Erol Aksöz

  2. Department of Chemistry, Biochemistry Division, Hacettepe University, P.K. 1, Samanpazari, 06242, Ankara, Turkey

    Adil Denizli

Authors
  1. Işık Perçin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Erol Aksöz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Adil Denizli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Adil Denizli.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Perçin, I., Aksöz, E. & Denizli, A. Gelatin-Immobilised Poly(hydroxyethyl methacrylate) Cryogel for Affinity Purification of Fibronectin. Appl Biochem Biotechnol 171, 352–365 (2013). https://doi.org/10.1007/s12010-013-0352-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-013-0352-6

Keywords

Search

Navigation