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Monitoring proteolysis of recombinant human interferon-γ during batch culture of Chinese hamster ovary cells

Goldman, Merlin H., James, David C., Ison, Andrew P., Bull, Alan T. (1997) Monitoring proteolysis of recombinant human interferon-γ during batch culture of Chinese hamster ovary cells. Cytotechnology, 23 (1-3). pp. 103-111. ISSN 0920-9069. (doi:10.1023/A:1007947130709) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1023/A:1007947130709

Abstract

Proteolytic cleavage of recombinant human interferon-? (IFN-?) expressed in Chinese hamster ovary (CHO) cells during batch fermentation has been monitored by mass spectrometric peptide mapping. IFN-? was purified from cell-free culture supernatant by immunoaffinity chromatography and cleaved by endoprotease Asp-N. Peptide fragments were resolved by reverse-phase HPLC and identified by a combination of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and automated N-terminal peptide sequencing. Using this approach, a peptide was identified as the C-terminal fragment of the IFN-? polypeptide. Analysis of this peptide by MS indicated that the recombinant IFN-? polypeptide secreted by CHO cells was truncated by at least ten amino acids, initially at Gln133-Met134. No full length (143 amino acids) polypeptide molecules were observed at any stages of the fermentation. Additional proteolytic cleavages at basic amino acids N-terminal of Gln133 occurred during the later stages of the culture resulting in a heterogeneous IFN-? polypeptide population with 'ragged' C-termini.

Item Type: Article
DOI/Identification number: 10.1023/A:1007947130709
Uncontrolled keywords: batch culture, Chinese hamster ovary, interferon-γ, mass spectrometry, proteolytic processing, serum-free cultivation
Subjects: Q Science > Q Science (General)
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: P. Ogbuji
Date Deposited: 29 May 2009 13:20 UTC
Last Modified: 01 Aug 2019 10:32 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/19602 (The current URI for this page, for reference purposes)
Bull, Alan T.: https://orcid.org/0000-0001-6223-7722
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