Recombinant DNA Technology and Characterization of Recombinant Alpha-2 Interferon

  • T. L. Nagabhushan
  • P. J. Leibowitz
Part of the Developments in Oncology book series (DION, volume 27)

Abstract

The strain of E. coli, KMAC-43, used for the large-scale production of alpha-2 interferon was engineered at Schering by Leibowitz and his co-workers. The alpha-2 interferon gene used in this construction was obtained from a cDNA clone, Hif-SN206, developed by Weissmann. The large-scale method developed in our laboratory for the purification of alpha-2 interferon from the bacterial extracts produces highly pure, crystalline alpha-2 interferon which is dissolved in 20 mM phosphate buffer at neutral pH to constitute bulk drug solution that is used in various formulations.

The protein has been thoroughly characterized using physico-chemical and biological methods. The average specific activity of clinical grade alpha-2 interfe­ron is 1.7 × 108 u/mgP. The purity of the product has been established by reversed phase HPLC (>98% pure), amino acid analysis, amino acid sequen­cing, circular dichroism spectrometry, ultracentrifugation, SDS-PAGE and 2-D gel electrophoresis. Details of these methods and results will be discussed.

Keywords

Fermentation HPLC Codon Cysteine Proline 

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Copyright information

© Martinus Nijhoff Publishers, Boston 1985

Authors and Affiliations

  • T. L. Nagabhushan
  • P. J. Leibowitz

There are no affiliations available

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