Characterization of DNA in cell culture supernatant by fluorescence-detection size-exclusion chromatography

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Characterization of DNA in cell culture supernatant by fluorescence-detection size-exclusion chromatography
Title:
Characterization of DNA in cell culture supernatant by fluorescence-detection size-exclusion chromatography
Journal Title:
Analytical and Bioanalytical Chemistry
Keywords:
Publication Date:
28 March 2015
Citation:
Tan L, Yeo V, Yang Y, Gagnon P. Characterization of DNA in cell culture supernatant by fluorescence-detection size-exclusion chromatography. Anal Bioanal Chem. 2015;407(14):4173‐4181. doi:10.1007/s00216-015-8639-9
Abstract:
A fluorescence-detection size-exclusion chromatography (FSEC) method was developed to characterize DNA in cell culture supernatant. Samples stained with Picogreen were fractionated by size-exclusion chromatography (SEC) and monitored simultaneously by UV absorbance and fluorescence. SEC provided a size-characterization capability absent from bulk fluorescent assays, and was also free from interference from other fluorescent and UV-absorbing small-molecule cell culture components. FSEC revealed that DNA in mammalian cell culture supernatant exists mostly in the form of nucleosomal arrays. FSEC combined with agarose electrophoresis revealed spontaneous degradation of DNA in mammalian cell culture supernatant over a 30 day period at 4 °C: from arrays containing up to ~40 nucleosomes, down to arrays containing three or fewer nucleosomes. It also detected nucleosomal DNA in wheat, soy, and yeast hydrolysates commonly used to enhance cell culture productivity.
License type:
PublisherCopyrights
Funding Info:
This research / project is supported by the Exploit Technologies Pte Ltd, A*STAR, under its ETPL GAP-Funded (Project No. ETPL/12-R15GAP-0009).
Description:
ISSN:
1618-2642
1618-2650
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