Info

aAfter each use, elution was carried out and then the gel was regenerated.

aAfter each use, elution was carried out and then the gel was regenerated.

33. In order to demonstrate the feasibility of reusing the support, several cycles of immobilization, elution of bound protein, regeneration of thiol-reactive adsorbent (TSI-gel), and reuse is performed. As high amounts of a model thiol-protein are required, bovine serum albumine-SH, provided with 4.0 moles of de novo thiol groups per mole of protein through chemical modification with SPDP, is selected for this purpose (Table 5).

References

1. Carlsson, J., Batista-Viera, F., and Rydén, L. (1998) Covalent chromatography. In: Protein Purification: Principles: High-Resolution Methods, and Applications, 2nd edition, Janson, J. C. and Rydén, L., eds,.. Wiley-VCH, New York, NY, pp. 343-373.

2. Carlsson, J., Axén, R., Brocklehurst, K., and Crook, E. (1974) Immobilization of urease by thiol-disulphide interchange with concomitant purification. Eur. J. Biochem. 44, 189-194.

3. Carlsson, J., Axén, R., and Unge, T. (1975) Reversible, covalent immobilization of enzymes by thiol-disulphide interchange. Eur. J. Biochem. 59, 567-572.

4. Oscarsson, S. and Porath, J. (1989) Covalent chromatography and salt-promoted thiophilic adsorption. Anal. Biochem. 176, 330-337.

5. Carlsson, J. and Batista-Viera, F. (1991) Solid phase disulfide oxides: a new approach to reversible immobilization and covalent chromatography of thiol compounds. Biotechnol. Appl. Biochem. 14, 114-120.

6. Batista-Viera, F., Barbieri, M., Ovsejevi, K., Manta, C., and Carlsson, J. (1991) A new method for reversible immobilization of thiol biomolecules based on solid phase bound thiolsulfonate groups. Appl. Biochem. Biotechnol. 31, 175-195.

7. Batista-Viera, F., Manta, C., and Carlsson, J. (1994) Solid-phase thiolsulfinates for the reversible immobilization of thiols. Appl. Biochem. Biotechnol. 44, 1-14.

8. Batista-Viera, F., Manta, C., and Carlsson, J. (1996) Covalent binding of thiols to thiolsulfinate- containing supports. Biotechnol. Appl. Biochem. 24, 231-239.

9. Axén, R., Drevin, H., and Carlsson, J.(1975) Preparation of modified agarose gels containing thiol groups. Acta Chem. Scand., B29, 471-474.

10. Ovsejevi, K., Brena, B., Batista-Viera, F., and Carlsson, J. (1995) Immobilization of E. coli P-galactosidase on thiolsulfonate agarose. Enzyme Microb. Technol. 17,151-156.

11. Ovsejevi, K., Grazú, V., and Batista-Viera, F. (1998) P-Galactosidase from Kluyveromices lactis immobilized on to thiolsulfinate/thiolsulfonate supports for lactose hydrolysis in milk and dairy by-products. Biotechnol. Tech. 12, 143-148.

12. Grazú, V., Ovsejevi, K., Cuadra, K., Betancor, L., Manta, C., and Batista-Viera, F. (2003) Solid phase reducing agents as alternative for reducing disulfide bonds in proteins. Appl. Biochem Biotechnol. 110, 23-32.

13. Brena, B., Ovsejevi, K., Luna, B., and Batista-Viera, F. (1993) Thiolation and reversible immobilization of sweet potato beta-amylase on thiolsulfonate-agar-ose. J. Mol. Catal. 84, 381-390.

14. Díaz, T., Stahl, U., Batista-Viera, F., and Carlsson, J. (1995) Reversible immobilization of chemically modified pullulanase. Biotechnol. Tech. 9, 533-538.

15. Persson, M., Bülow, L., and Mosbach, K. (1990) Purification and site-specific immobilization of genetically engineered glucose dehydrogenase on Thiopropyl-Sepharose. FEBS Lett. 270, 41-44.

16. Irazoqui, G., Villarino, A., Batista-Viera, F., and Brena, B. (2002) Generating favorable nano-environments for thermal and solvent stabilization of immobilized P-galactosidase. Biotechnol. Bioeng. 77, 430-434.

17. Brocklehurst, K., Carlsson, J., Kierstan, M., and Crook, E. (1973) Covalent chromatography. Preparation of fully active papain from dried papaya latex. Biochem. J. 133, 573-584.

18. Worthington, V. (1993) P-Galactosidase . In Worthington Enzyme Manual, Freehold, NJ, p.179.

19. MERCK Schuchardt technical information (1988) A versatile new peroxyacid. Magnesium monoperoxyphthalate. MS Info 88-1.

20. Ali, M. and Stevens, W.(1997) A facile and selective procedure for oxidation of sulfides and sulfoxides on silica gel supported magnesium monoperoxyphthalate (MMPP) in dichloromethane. Synthesis 7, 764-768.

21. Ellman, G. L. (1958) A colorimetric method for determining low concentrations of mercaptans. Arch. Biochem. Biophys. 74, 443-450.

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