Rapid and reliable quantification of reovirus type 3 by high performance liquid chromatography during manufacturing of Reolysin

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DOIResolve DOI: http://doi.org/10.1016/j.jpba.2008.05.038
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleJournal of Pharmaceutical and Biomedical Analysis
ISSN0731-7085
Volume48
Issue3
Pages598605; # of pages: 8
Subjectreovirus; quantification; high performance liquid chromatography
AbstractReolysin, a human reovirus type 3, is being evaluated in the clinic as an oncolytic therapy for various types of cancer. To facilitate the optimization and scale-up of the current process, a high performance liquid chromatography (HPLC) method has been developed that is rapid, specific and reliable for the quantification of reovirus type 3 particles. Using an anion-exchange column, the intact virus eluted from the contaminants in 9.78 min at 350 mM NaCl in 50mM HEPES, pH 7.10 in a total analysis time of 25 min. The virus demonstrated a homogenous peak with no co-elution of other compounds as analyzed by photodiode array analysis. The HPLC method facilitated the optimization of the purification process which resulted in the improvement of both total and infectious particle recovery and contributed to the successful scale-up of the process at the 20 L, 40 L and 100 L production scale. The method is suitable for the analysis of crude virus supernatants, crude lysates, semi-purified and purified preparations and therefore is an ideal monitoring tool during process development and scale-up.
Publication date
PublisherElsevier
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedYes
NRC number47826
NPARC number21268261
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Record identifierc4c7b023-cd79-4512-9af5-7221d151e967
Record created2013-06-11
Record modified2016-05-09
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