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Identification of cellular changes associated with increased production of human follicle stimulating hormone in a recombinant Chinese hamster ovary cell line

A proteomics approach was used to identify proteins potentially implicated in the cellular response concurrent with elevated production levels of human follicle stimulating hormone in a recombinant Chinese hamster ovary cell line (Darren cells), using zinc and sodium butyrate in the production media to increase expression. To this end, 2-dimensional gel electrophoresis (2-DGE) was utilized. Firstly, several aspects of 2-DGE were developed for this investigation. Gel drying conditions were optimized, and a glycine-free blotting method is described which achieved greater efficiency in rapid transfer of proteins than those previously described. Next, hFSH expression was characterized in Darren cells. An ELISA developed for this investigation examined intracellular (expression) and extracellular (secretion) of hFSH during increased expression. These results show a disproportionate increase in intracellular hFSH (188%) expression above extracellular hFSH (41%).

Identiferoai:union.ndltd.org:ADTP/212654
Date January 2008
CreatorsMisztal, David Richard, Biotechnology & Biomolecular Sciences, Faculty of Science, UNSW
PublisherPublisher:University of New South Wales. Biotechnology & Biomolecular Sciences
Source SetsAustraliasian Digital Theses Program
LanguageEnglish
Detected LanguageEnglish
Rightshttp://unsworks.unsw.edu.au/copyright, http://unsworks.unsw.edu.au/copyright

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