Loading [MathJax]/extensions/tex2jax.js

Production of interferon-α in high cell density cultures of recombinant Escherichia coli and its single step purification from refolded inclusion body proteins

Research output: Contribution to journalArticleResearchpeer review

Authors

  • K. R. Babu
  • S. Swaminathan
  • S. Marten
  • N. Khanna
  • U. Rinas

External Research Organisations

  • International Centre for Genetic Engineering and Biotechnology India
  • Helmholtz Centre for Infection Research (HZI)

Details

Original languageEnglish
Pages (from-to)655-660
Number of pages6
JournalApplied Microbiology and Biotechnology
Volume53
Issue number6
Publication statusPublished - Jun 2000
Externally publishedYes

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

Cite this

Production of interferon-α in high cell density cultures of recombinant Escherichia coli and its single step purification from refolded inclusion body proteins. / Babu, K. R.; Swaminathan, S.; Marten, S. et al.
In: Applied Microbiology and Biotechnology, Vol. 53, No. 6, 06.2000, p. 655-660.

Research output: Contribution to journalArticleResearchpeer review

Download
@article{6b7abbd807d6475ea5127731739cfc47,
title = "Production of interferon-α in high cell density cultures of recombinant Escherichia coli and its single step purification from refolded inclusion body proteins",
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.",
author = "Babu, {K. R.} and S. Swaminathan and S. Marten and N. Khanna and U. Rinas",
year = "2000",
month = jun,
doi = "10.1007/s002530000318",
language = "English",
volume = "53",
pages = "655--660",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "6",

}

Download

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 -