A Flow Cytometry-Based Method to Determine the Titer of Adenoviruses Expressing an Extraneous Gene
| Autor: | Kanae Abe, Yasuo Urata, Natsuki Yanase, Hiroyuki Mizuguchi, Akikazu Asada, Hideki Hayakawa, Fuminori Sakurai |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Oncolytic adenovirus Adenoviridae Infections Green Fluorescent Proteins Pharmaceutical Science Biology Flow cytometry Green fluorescent protein Adenoviridae 03 medical and health sciences 0302 clinical medicine Cell Line Tumor medicine Humans Pharmacology medicine.diagnostic_test Infectious dose General Medicine Viral Load Flow Cytometry Virology Oncolytic virus Titer Oncolytic Viruses 030104 developmental biology Cell culture 030220 oncology & carcinogenesis Microorganisms Genetically-Modified Viral load HeLa Cells |
| Zdroj: | Biologicalpharmaceutical bulletin. 41(10) |
| ISSN: | 1347-5215 |
| Popis: | In recent times, oncolytic viruses expressing an extraneous gene have attracted great interest; in fact, they have been engaged in multiple applications, such as medicine for cancer. Our group made an oncolytic adenovirus, namely, OBP-301, for use in treating solid cancers and press clinical trial to get approval for a pharmaceutical product. In this study, we applied a flow cytometry-based method to determine the titer of adenoviruses expressing an extraneous gene as well as assess their quality. We considered using the green fluorescent protein (GFP)50 titer as a measure of viral quality. The GFP50 titer (GFP50/mL) is the viral load required to render the HeLa S3 cell line 50% GFP-positive by analysing flow cytometry data. We measured the GFP50 titers for three types of recombinant adenoviruses (OBP-401, OBP-1101, and OBP-1106). We compared GFP50/mL and tissue culture infectious dose (TCID50/mL), a conventional titration index, and found that these titers showed a linear correlation, with a correlation coefficient of >0.9. Moreover, GFP50/mL showed high repetitive accuracy. We expect this flow cytometry-based method to be useful in case of clinically relevant viruses expressing an extraneous gene, in particular, to control viral quality. |
| Databáze: | OpenAIRE |
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