Details
Original language | English |
---|---|
Pages (from-to) | 655-660 |
Number of pages | 6 |
Journal | Applied Microbiology and Biotechnology |
Volume | 53 |
Issue number | 6 |
Publication status | Published - Jun 2000 |
Externally published | Yes |
Abstract
Escherichia coli TG1 transformed with a temperature-regulated interferon-α expression vector was grown to high cell density in defined medium containing glucose as the sole carbon and energy source, utilizing a simple fed-batch process. Feeding was carried out to achieve an exponential increase in biomass at growth rates which minimized acetate production. Thermal induction of such high cell density cultures resulted in the production of ~4 g interferon-α/l culture broth. Interferon-α was produced exclusively in the form of insoluble inclusion bodies and was solubilized under denaturing conditions, refolded in the presence of arginine and purified to near homogeneity, utilizing single-step ion-exchange chromatography on Q-Sepharose. The yield of purified interferon-α was ~300 mg/l with respect to the original high cell density culture broth (overall yield of ~7.5% active interferon-α). The purified recombinant interferon-α was found by different criteria to be predominantly monomeric and possessed a specific bioactivity of ~2.5 x 108 IU/mg based on viral cytopathic assay.
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Applied Microbiology and Biotechnology, Vol. 53, No. 6, 06.2000, p. 655-660.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Production of interferon-α in high cell density cultures of recombinant Escherichia coli and its single step purification from refolded inclusion body proteins
AU - Babu, K. R.
AU - Swaminathan, S.
AU - Marten, S.
AU - Khanna, N.
AU - Rinas, U.
PY - 2000/6
Y1 - 2000/6
N2 - Escherichia coli TG1 transformed with a temperature-regulated interferon-α expression vector was grown to high cell density in defined medium containing glucose as the sole carbon and energy source, utilizing a simple fed-batch process. Feeding was carried out to achieve an exponential increase in biomass at growth rates which minimized acetate production. Thermal induction of such high cell density cultures resulted in the production of ~4 g interferon-α/l culture broth. Interferon-α was produced exclusively in the form of insoluble inclusion bodies and was solubilized under denaturing conditions, refolded in the presence of arginine and purified to near homogeneity, utilizing single-step ion-exchange chromatography on Q-Sepharose. The yield of purified interferon-α was ~300 mg/l with respect to the original high cell density culture broth (overall yield of ~7.5% active interferon-α). The purified recombinant interferon-α was found by different criteria to be predominantly monomeric and possessed a specific bioactivity of ~2.5 x 108 IU/mg based on viral cytopathic assay.
AB - Escherichia coli TG1 transformed with a temperature-regulated interferon-α expression vector was grown to high cell density in defined medium containing glucose as the sole carbon and energy source, utilizing a simple fed-batch process. Feeding was carried out to achieve an exponential increase in biomass at growth rates which minimized acetate production. Thermal induction of such high cell density cultures resulted in the production of ~4 g interferon-α/l culture broth. Interferon-α was produced exclusively in the form of insoluble inclusion bodies and was solubilized under denaturing conditions, refolded in the presence of arginine and purified to near homogeneity, utilizing single-step ion-exchange chromatography on Q-Sepharose. The yield of purified interferon-α was ~300 mg/l with respect to the original high cell density culture broth (overall yield of ~7.5% active interferon-α). The purified recombinant interferon-α was found by different criteria to be predominantly monomeric and possessed a specific bioactivity of ~2.5 x 108 IU/mg based on viral cytopathic assay.
UR - http://www.scopus.com/inward/record.url?scp=0033920414&partnerID=8YFLogxK
U2 - 10.1007/s002530000318
DO - 10.1007/s002530000318
M3 - Article
C2 - 10919322
AN - SCOPUS:0033920414
VL - 53
SP - 655
EP - 660
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 6
ER -