Goldman, Merlin H. and James, David C. and Ison, Andrew P. and 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 . (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)
Proteolytic cleavage of recombinant human interferon-γ (IFN-γ) expressed in Chinese hamster ovary (CHO) cells during batch fermentation has been monitored by mass spectrometric peptide mapping. IFN-γ 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-γ polypeptide. Analysis of this peptide by MS indicated that the recombinant IFN-γ 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-γ polypeptide population with 'ragged' C-termini.
|Subjects:||Q Science > Q Science (General)|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||P. Ogbuji|
|Date Deposited:||29 May 2009 13:20|
|Last Modified:||10 Jul 2014 14:19|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/19602 (The current URI for this page, for reference purposes)|