Production and purification of human menin from Drosophila melanogaster S2 cells using stirred tank reactor (original) (raw)

Abstract

A process was developed for producing human menin from transformed Drosophila Schneider 2 cells. Protein expression was achieved after inducing the metallothionein promoter by adding copper sulfate to cells growing in suspension in a stirred-tank reactor. Experiments in shake flasks showed that the production of menin was improved when the induction was conducted late in the exponential phase of cell growth at a concentration of 1–2 × 107 cells ml-1, with a copper concentration of 0.2 mM for no more than 24 h. This observation was confirmed by experiments in bench-scale fermentors. Subsequently, a pilot-scale fermentation yielded 1 mg l-1 culture of purified menin.

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References

• Bunch TA, Grinblat Y and Goldstain, LSB (1988) Characterization and use of the Drosophila metallothionein promoter in cultured Drosophila melanogaster cells. Nucleic Acids Res 16: 1043–1061.
  PubMed CAS Google Scholar
• Box GEP, Hunter WG and Hunter JS (1987) Statistics for Experiments (pp. 310–351) John Wiley & Sons, New York.
  Google Scholar
• Castillo AJ, Fernandez A, Boggiano T, Pugeaud P and Marison IW (1998) Study of different cell culture conditions for the production of a ‘reshaped’ MAb in NSO Cells. In: Merten O-W et al. (eds.) New Developments and New Applications in Animal Cell Technology (pp. 51–53) Kluwer Academic Publishers, Dordrecht.
  Google Scholar
• Chandrasekharappa SC, Guru SC, Manickam P, Olufemi SE, Collins FS, Emmert-Buck MR, Debelenko LV, Zhuang Z, Lubensky IA, Liotta LA, Crabtree JS, Wang Y, Roe BA, Weisemann J, Boguski MS, Agarwal SK, Kester MB, Kim YS, Heppner, C, Dong Q, Spiegel AM, Burns AL and Marx SJ (1997) Positional cloning of the gene for multiple endocrine-neoplasia-Type 1. Science 276: 404–407.
  Article PubMed CAS Google Scholar
• Deml L, Wolf H and Wagner R (1999) High level expression of hepatitis B virus surface antigen in stably transfected Drosophila Schneider-2 cells. J Virol Methods 79: 191–203.
  Article PubMed CAS Google Scholar
• Guru SC, Goldsmith PK, Burns AL, Marx SJ, Spiegel AM, Collins FS and Chandrasekharappa SC (1998) Menin, the product of the MEN1 gene, is a nuclear protein. Proc Nat Acad Sci USA 95: 1630–1634.
  Article PubMed CAS Google Scholar
• Hansen BE, Andersson EC, Madsen LS, Engberg J, Sondergaard L, Svejgaard A and Fugger L (1998) Functional characterization of HLA-DRA1*0101/DRB1*0401 molecules expressed in Drosophila melanogaster cells. Tissue Antigens 51: 119–128.
  Article PubMed CAS Google Scholar
• Johansen H, van der Straten, Sweet R, Otto E, Maroni G and Rosenberg M (1989) Regulated expression at high copy number allows production of a growth-inhibitory oncogene product in Drosophila Schneider cells. Genes Dev 3: 882–889.
  PubMed CAS Google Scholar
• Li B, Tsing S, Kosaka AH, Nguyen B, Osen EG, Bach C, Chan H and Barnett J (1996) Expression of human dopaminehydroxylase in Drosophila Schneider 2 cells. Biochem J 313: 57–64.
  PubMed CAS Google Scholar
• Marx SJ, Agarwal SK, Kester MB, Heppner C, Kim YS, Skarulis MC, James LA, Goldsmith PK, Saggar SK, Park SY, Spiegel AM, Burns AL, Debelenko LV, Zhuang Z, Lubensky IA, Liotta LA, Emmert-Buck MR, Guru SC, Manickam P, Crabtree J, Erdos MR, Collins FS and Chandrasekharappa SC (1999) Multiple endocrine neoplasia type 1: Clinical and genetic features of the hereditary endocrine neoplasias. Recent Prog Horm Res 54: 397–439.
  PubMed CAS Google Scholar
• Nilsen SL and Castellino FJ (1999) Expression of human plasminogen in Drosophila Schneider S2 cells. Protein Expr Purif 16: 136–143.
  Article PubMed CAS Google Scholar
• Olsen MK, Rockenbach SK, Fischer HD, Hoogerheide JG and Tomich CSC (1992) Stable production of an analog of human tissue plasminogen-activator from cultured Drosophila cells. Cytotechnology 10: 157–167.
  Article PubMed CAS Google Scholar
• Park JH, Lee JM and Chung IS (1999) Production of recombinant endostatin from stably tranformed Drosophila melanogaster S 2 cells. Biotechnol Lett 21: 729–733.
  Article CAS Google Scholar
• Schneider I (1972) Cell lines derived from late embryonic stages of Drosophila melanogaster. J Embryol Exp Morphol 27: 353–365.
  PubMed CAS Google Scholar
• Sondergaard L (1996) Drosophila cells can be grown to high cell density in a bioreactor. Biotechnol Tech 10: 161–166.
  Article CAS Google Scholar
• Watanabe S, Shimazaki K, Bollen A and Moguilevsky N (1998) Characterization of recombinant bovine lactoperoxidase produced by CHO cells. In: Ikura K et al. (eds) Animal Cell Technology: Challenges for the 21st Century (pp. 93–97), Kluwer Academic Publishers, Dordrecht.
  Google Scholar

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Authors and Affiliations

1. Biotechnology Unit, NIDDK, NIH, Bethesda, MD, 20892, U.S.A.
   M. A. Valle
2. Center for Agriculture Biotechnology, University of Maryland, U.S.A.
   M. A. Valle
3. Metabolic Diseases Branch, NIDDK, NIH, U.S.A.
   M. B. Kester, A. L. Burns, S. J. Marx & A. M. Spiegel
4. Biotechnology Unit, NIDDK, NIH, Bethesda, MD, 20892, U.S.A.
   J. Shiloach

Authors

1. M. A. Valle
2. M. B. Kester
3. A. L. Burns
4. S. J. Marx
5. A. M. Spiegel
6. J. Shiloach

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Correspondence toJ. Shiloach.

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Valle, M.A., Kester, M.B., Burns, A.L. et al. Production and purification of human menin from Drosophila melanogaster S2 cells using stirred tank reactor.Cytotechnology 35, 127–135 (2001). https://doi.org/10.1023/A:1017586523710

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• Issue date: March 2001
• DOI: https://doi.org/10.1023/A:1017586523710