May 8-13, 2016
Chinese Hamster Ovary (CHO) cell lines are common hosts for the production of biotherapeutic proteins. Achieving high level of specific protein production by CHO cell lines remains a challenge. In order to address this issue, we are incorporating lipidomic analyses to study the role of lipids played in CHO-S cells.
In our study, we have applied chromatography (TLC) methods for lipid analysis in terms of lipid polarity. For polar lipids, 2-D HPTLC (2-dimensional high performance TLC) was used instead of conventional 1D- TLC by virtue of its high separation capacity. The eluting solvent system was optimized for the 1st and 2nd dimension, respectively. Neutral lipids were separated on 1-D HPTLC with the optimal elution solvent of hexane-diethyl ether-acetic acid. The lipid spots on the TLC plates were stained by 0.2% of 2,7-dichlorofluorescein dissolved in ethanol solution and illuminated with UV. Multiple lipid standards were also run to correctly identify the lipid spots and the fluorescence of lipid spots was semi-quantitatively measured with ImageJ.
By optimization of TLC conditions, the lipids of CHO-S cell line were separated successfully and the lipid contents were semi-quantified. From neutral lipids result, we observed high level of certain lipids in CHO-S cell lines. We will further investigate which lipid play a key role in various cell processes.
Yue Zhang, Deniz Baycin, Michael Bowen, and Michael Betenbaugh, "Lipidomic analysis to enhance the understanding of Chinese Hamster ovary cells" in "Cell Culture Engineering XV", Robert Kiss, Genentech Sarah Harcum, Clemson University Jeff Chalmers, Ohio State University Eds, ECI Symposium Series, (2016). http://dc.engconfintl.org/cellculture_xv/174