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Solubilization and Refolding with Simultaneous Purification of Recombinant Human Stem Cell Factor

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Abstract

Recombinant human stem cell factor (rhSCF) was solubilized and renatured from inclusion bodies expressed in Escherichia coli. The effect of both pH and urea on the solubilization of rhSCF inclusion bodies was investigated; the results indicate that the solubilization of rhSCF inclusion bodies was significantly influenced by the pH of the solution employed, and low concentration of urea can drastically improve the solubilization of rhSCF when solubilized by high pH solution. The solubilized rhSCF can be easily refolded with simultaneous purification by ion exchange chromatography (IEC), with a specific activity of 7.8 × 105 IU·mg−1, a purity of 96.3%, and a mass recovery of 43.0%. The presented experimental results show that rhSCF solubilized by high pH solution containing low concentration of urea is easier to be renatured than that solubilized by high concentration of urea, and the IEC refolding method was more efficient than dilution refolding and dialysis refolding for rhSCF. It may have a great potential for large-scale production of rhSCF.

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References

  1. Galli, S. J., Zsebo, K. M., & Geissler, E. N. (1994). Advances in Immunology, 55, 1–96.

    Article  CAS  Google Scholar 

  2. Broudy, V. C. (1997). Blood, 90, 1345–1364.

    CAS  Google Scholar 

  3. Lu, H. S., Clogston, C. L., Wypych, J., Fausset, P. R., Lauren, S., Mendiaz, E. A., et al. (1991). Journal of Biological Chemistry, 266, 8102–8107.

    CAS  Google Scholar 

  4. Langley, K. E., Wypych, J., Mendiaz, E. A., Clogston, C. L., Parker, V. P., Farrar, D. H., et al. (1992). Archives of Biochemistry and Biophysics, 295, 21–28.

    Article  CAS  Google Scholar 

  5. Hsu, Y. R., Wu, G. M., Mendiaz, E. A., Syed, R., Wypych, J., Toso, R., et al. (1997). Journal of Biological Chemistry, 272, 6406–6415.

    Article  CAS  Google Scholar 

  6. Wang, L. L., Geng, X. D., & Han, H. (2004). China Journal of Molecular Immunology, 20, 402–405.

    CAS  Google Scholar 

  7. Wu, J., Gong, X., Chang, S. H., Zhao, Z. H., Zuo, C. L., & Ma, Q. J. (2003). Chinese Journal of Biotechnology, 19, 698–704.

    CAS  Google Scholar 

  8. Wu, J., Gong, X., Chang, S. H., Zhao, Z. H., & Ma, Q. J. (2004). Chinese Journal of Biochemistry and Molecular Biology, 20, 821–826.

    CAS  Google Scholar 

  9. Tsumoto, K., Ejima, D., Kumagai, I., et al. (2003). Protein Expression and Purification, 28, 1–8.

    Article  CAS  Google Scholar 

  10. Khan, R. H., AppaRao, K. B. C., Eshwari, A. N. S., et al. (1998). Biotechnology Progress, 14, 722–728.

    Article  CAS  Google Scholar 

  11. Shin, N. K., Kim, D. Y., Shin, C. S., et al. (1998). Journal of Biotechnology, 62, 143–151.

    Article  CAS  Google Scholar 

  12. Hartman, J. R., Mendelovitz, S., Gorecki, M. (1999). US Patent 6,001,604.

  13. Geng, X. D., & Wang, C. Z. (2007). Journal of Chromatography B, 849, 69–80.

    Article  CAS  Google Scholar 

  14. Wang, C. Z., Geng, X. D., Wang, D. W., & Tian, B. (2004). Journal of Chromatography B, 806, 185–190.

    Article  CAS  Google Scholar 

  15. Wang, C. Z., Wang, L. L., & Geng, X. D. (2006). Journal of Liquid Chromatography and Related Techniques, 29, 203–217.

    Article  CAS  Google Scholar 

  16. Jungbauer, A., Kaar, W., & Schlegl, R. (2004). Current Opinion in Biotechnology, 15, 487–494.

    Article  CAS  Google Scholar 

  17. Li, M., Su, Z. G., & Janson, J. C. (2004). Protein Expression and Purification, 33, 1–10.

    Article  CAS  Google Scholar 

  18. Geng, X. D., Bai, Q., & Wang, C. Z. (2006). Protein folding liquid chromatography. Beijing: Science in China.

    Google Scholar 

  19. Liu, X. Q., Yang, X. Q., Xie, F. H., et al. (2007). Protein Expression and Purification, 51, 179–186.

    Article  CAS  Google Scholar 

  20. Liu, H. N., Wang, Y., Gong, B. L., & Geng, X. D. (2005). Acta Chimica Sinica, 63, 597–602.

    CAS  Google Scholar 

  21. Franzmann, T. M. (2006). International Journal of Biological Macromolecules, 39, 104–110.

    Article  CAS  Google Scholar 

  22. Langenhof, M., Leong, S. S. J., Pattenden, L. K., et al. (2005). Journal of Chromatography A, 1069, 195–201.

    Article  CAS  Google Scholar 

  23. Li, M., Zhang, G. F., & Su, Z. G. (2002). Journal of Chromatography A, 959, 113–120.

    Article  CAS  Google Scholar 

  24. Creighton, T. E. (1990). US Patent 4,977,248.

  25. Stempfer, G., Holl-Neugebauer, B., & Rudolph, R. (1996). Nature Biotechnology, 14, 329–334.

    Article  CAS  Google Scholar 

  26. Wang, Y., Gong, B. L., & Geng, X. D. (2003). Chinese Chemical Letters, 14, 828–831.

    CAS  Google Scholar 

  27. Wang, C. Z., Wang L. L., & Geng X. D. (2007). Biomedical Chromatography, 21, 1291–1296.

    Article  CAS  Google Scholar 

  28. Machold, C., Schlegl, R., Buchinger, W., & Jungbauer, A. (2005). Journal of Chromatography A, 1080, 29–42.

    Article  CAS  Google Scholar 

  29. Wang, J. Z., Zhao, Y., Chen, G. Q., & Rao, C. M. (2001). China Journal of Cancer Biotherapy, 8, 294–296.

    Google Scholar 

  30. Patra, A. K., Mukhopadhyay, R., Mukhija, R., et al. (2000). Protein Expression and Purification, 18, 182–192.

    Article  CAS  Google Scholar 

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Acknowledgment

This work was supported by the National Natural Science Foundation in China (no. 20705028) and the Foundation of Key Laboratory of Modern Separation Science in Shaanxi Province (no. 05JS61).

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Authors and Affiliations

  1. Institute of Modern Separation Science, Department of Chemistry, Northwest University, 229 Tai Bai North Road, Xi’an, 710069, People’s Republic of China

    Chaozhan Wang, Jiahua Liu, Lili Wang & Xindu Geng

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  1. Chaozhan Wang
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  2. Jiahua Liu
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  3. Lili Wang
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  4. Xindu Geng
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Correspondence to Chaozhan Wang.

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Wang, C., Liu, J., Wang, L. et al. Solubilization and Refolding with Simultaneous Purification of Recombinant Human Stem Cell Factor. Appl Biochem Biotechnol 144, 181–189 (2008). https://doi.org/10.1007/s12010-007-8112-0

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  • DOI: https://doi.org/10.1007/s12010-007-8112-0

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