Production and purification of human menin from Drosophila melanogaster S2 cells using stirred tank reactor |
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Authors: | M. A. Valle M. B. Kester A. L. Burns S. J. Marx A. M. Spiegel J. Shiloach |
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Affiliation: | (1) Biotechnology Unit, NIDDK, NIH, Bethesda, MD, 20892, U.S.A.;(2) Center for Agriculture Biotechnology, University of Maryland, U.S.A.;(3) Metabolic Diseases Branch, NIDDK, NIH, U.S.A.;(4) Biotechnology Unit, NIDDK, NIH, Bethesda, MD, 20892, U.S.A. |
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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. This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | Drosophila Schneider menin |
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