İlker KOÇ, Nilay BERELİ, Gözde BAYDEMİR, Igor Yu GALAEV, Adil DENİZLİ

Synthesis and Characterization of Macroporous Poly Acrylamide-Methacrylamido Histidine Cryogels and Their Use in Antibody Purification

Poly acrylamide-methacrylamidohistidine [poly AAm-MAH ] cryogel was prepared by bulk polymerization whichproceeds in aqueous solution of monomers frozen inside a syringe column cryo-polymerization . After thawing, themonolithic cryogel contains a continuous polymeric matrix having interconnected pores of 10-100 µm size. Poly AAmMAH cryogel was characterized by swelling studies, FTIR and scanning electron microscopy. The equilibrium swellingdegree of the poly AAm-MAH monolithic cryogel was 6.97 g H2O/g cryogel. Poly AAm-MAH cryogel was used in theadsorption/desorption of immunoglobulin G IgG from aqueous solutions and human plasma. The non-specificadsorption of IgG was very low 0.22 mg/g . The maximum amount of IgG adsorption from aqueous solution in HEPESbuffer was 32.879 mg/g at pH 7.4. Higher IgG adsorption value was obtained from human plasma up to 100 mg/g .Desorption of IgG from poly AAm-MAH cryogel was obtained using 20 mM Acetate buffer containing 1.0 M NaCl atpH 4. It was observed that IgG could be repeatedly adsorbed and desorbed with poly AAm-MAH cryogel withoutsignificant loss in the adsorption capacity

Keywords:

Affinity chromatography, cryogel, Protein Purification, IgG,

PDF

___

1. V.I. Lozinsky, F.M. Plieva, I.Y. Galaev, B. Mattiasson, Bioseparation 10 (2002) 163.
2. V.I. Lozinsky, I.Y. Galaev, F.M. Plieva, I.N. Savina, H. Jungvid, B. Mattiasson, TRENDS in Biotechnol., 21 (2003) 445.
3. P. Arvidsson, F.M. Plieva, V.I. Lozinsky, I.Y. Galaev, B. Mattiasson, J. Chromatogr. A 986 (2003) 275.
4. P. Arvidsson, F.M. Plieva, I.N. Savina, V.I. Lozinsky, S. Fexby, L. Bulow, I.Y. Galaev, B. Mattiasson, J. Chromatogr. A, 977 (2002) 27.
5. F. Svec, Electrophoresis, 27 (2006) 947.
6. L. Uzun, R. Say, A. Denizli, React. Funct. Polym. 64 (2005) 93.
7. C. Babac, H. Yavuz, I.Y. Galaev, E. Pişkin, A. Denizli, React. & Functl. Polym. 66 (2006) 163.
8. Guerrier, L., Girot, P., Schwartz, W., Boschetti, E., Bioseparation, 9 (2000) 211.
9. G. Tishchenko, J. Dybal, K. Meszarosova, Z. Sedlakova, M. Bleha, J. Chromatogr. A, 954 (2002) 115.
10. V.I. Muronetz, T. Korpela, J. Chromatogr. B, 790 (2003) 53.
11. T. Burnouf, M. Radosevich, J. Biochem. Biophys. Methods, 49 (2001) 575.
12. Tejeda-Mansir, R. Espinoza, R.M. Montesinos, R. Guzman, Bioproc. Eng., 17 (1997) 39.
13. A. Denizli, J.Chromatogr.B, 668 (1995) 13.
14. P. Füglistaller, J. Immunol. Methods 124 (1989) 171.
15. Denizli, Y. Arica, J. Biomater. Sci. Polym. Ed. 11 (2000) 367.
16. O. Pitiot, C. Legallais, L. Darnige, M.A. Vijayalakshmi, J. Membr. Sci. 166 (2000) 221.
17. O. Pitiot, L. Folley, M.A. Vijayalakshmi, J. Chromatogr. B, 758 (2001) 163.
18. Huang, P.Y., and Carbonell, R.G., Biotechnol. Bioeng., 63 (1999) 633-641.
19. M.A. Vijayalakshmi, Trends Biotechnol. 7 (1989) 71.
20. K. Haupt, M.A. Vijayalakshmi, J. Chromatogr. 644 (1993) 289.
21. C.I. Alvarez, M.C. Strumia, H.E. Bertorello, React. Functl. Polym., 34 (1997) 103.
22. A. Legallais, F.B. Anspach, S.M.A. Bueno, K. Haupt, M.A. Vijayalakshmi, J. Chromatogr. B, 691 (1997) 33.
23. K. Haupt, S.M.A. Bueno, M.A. Vijayalakshmi, J. Chromatogr. B, 674 (1995) 13.
24. A. El-Kak, S. Manjini, M.A. Vijayalakshmi, J. Chromatogr. B, 604 (1992) 29.
25. S.M.A. Bueno, K. Haupt, M.A. Vijayalakshmi, J. Chromatogr. B, 66 (1995) 57.
26. E. Klein, E. Eichholtz, D.H. Yeager, J. Membrane Sci., 95 (1994) 199.
27. M. Kim, K. Saito, S. Furusaki, S., Sato, T., Sugo, T., Ishigaki, I., 1991, J. Chromatogr., 585, 45.
28. D. Müller-Schulte, S. Manjini, M.A. Vijayalakshmi, J. Chromatogr., 539 (1991) 307.
29. A. Denizli and E. Pişkin, J. Chromatogr. B, 666 (1995) 215-222.
30. P. Langlotz, K.H. Kroner, J. Chromatogr., 591 (1992) 107.
31. O.P. Dancette, J.L. Taboureau, E. Tournier, C. Charcosset, P. Blond, J. Chromatogr. B., 723 (1999) 61.
32. S.F. Teng, K. Sproule, A. Husain, C.R. Lowe, J. Chromatogr. B., 740 (2000) 1.
33. S. Özkara, H. Yavuz, S. Patir, Y. Arica, A. Denizli, Sep. Sci. Technol., 37 (2002) 717.
34. S.F. Teng, K. Sproule, A. Hussain, C.R. Lowe, J. Mol. Recog., 12 (1999) 67.
35. G. Fassina, A. Verdoliva, G. Palombo, M. Ruvo, G. Cassani, J. Mol. Recog. 1998, 11, 128.