دورية أكاديمية
Development of an HSV-1 production process involving serum-reduced culturing and bead-to-bead transfer.
| العنوان: | Development of an HSV-1 production process involving serum-reduced culturing and bead-to-bead transfer. |
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| المؤلفون: | Wang H; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Hu X; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Zhang M; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Yang L; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Xu Y; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Gu X; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China., Jiang J; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China. jiang_junjun@wuxibiologics.com., Hu W; Microbial and Viral Platforms (MVP), WuXi Biologics, 291 Fucheng Road, Hangzhou, 311106, China. hu_weiwei0501@wuxibiologics.com. |
| المصدر: | Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2024 Jun 19; Vol. 108 (1), pp. 383. Date of Electronic Publication: 2024 Jun 19. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer International Country of Publication: Germany NLM ID: 8406612 Publication Model: Electronic Cited Medium: Internet ISSN: 1432-0614 (Electronic) Linking ISSN: 01757598 NLM ISO Abbreviation: Appl Microbiol Biotechnol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Berlin ; New York : Springer International, c1984- |
| مواضيع طبية MeSH: | Herpesvirus 1, Human*/growth & development , Bioreactors*/virology , Culture Media*/chemistry , Virus Cultivation*/methods, Chlorocebus aethiops ; Vero Cells ; Animals |
| مستخلص: | Herpes simplex virus type 1 (HSV-1) plays an important role in the field of gene therapy and viral vaccines, especially as an oncolytic virus. However, the mass production of HSV-1 viral vectors remains a challenge in the industry. In this study, a microcarrier-mediated serum-reduced medium culture was used to improve the bioprocess of HSV-1 production and increase HSV-1 yields. The composition of the culture media, which included a basal medium, serum concentration, and glutamine additive, was optimized. The process was successfully conducted in a 1 L bioreactor, and virus production was threefold greater than that of conventional processes with a 10% serum medium. The bead-to-bead transfer process was also developed to further increase scalability. In spinner flasks, the detachment rate increased from 49.4 to 80.6% when combined agitation was performed during digestion; the overall recovery proportion increased from 37.9 to 71.1% after the operational steps were optimized. Specifically, microcarrier loss was reduced during aspiration and transfer, and microcarriers and detached cells were separated with filters. Comparable cell growth was achieved with the baseline process using 2D culture as the inoculum by exchanging the subculture medium. To increase virus production after bead-to-bead transfer, critical parameters, including shear stress during digestion, TrypLE and EDTA concentrations in the subculture, and the CCI, were identified from 47 parameters via correlation analysis and principal component analysis. The optimized bead-to-bead transfer process achieved an average of 90.4% overall recovery and comparable virus production compared to that of the baseline process. This study is the first to report the optimization of HSV-1 production in Vero cells cultured on microcarriers in serum-reduced medium after bead-to-bead transfer. KEY POINTS: • An HSV-1 production process was developed that involves culturing in serum-reduced medium, and this process achieved threefold greater virus production than that of traditional processes. • An indirect bead-to-bead transfer process was developed with over 90% recovery yield in bioreactors. • HSV-1 production after bead-to-bead transfer was optimized and was comparable to that achieved with 2D culture as inoculum. (© 2024. The Author(s).) |
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| فهرسة مساهمة: | Keywords: Bead-to-bead transfer; Herpes simplex virus type 1; Microcarriers; Serum-reduced culturing; Stirred-tank bioreactor; Vero cells |
| المشرفين على المادة: | 0 (Culture Media) |
| تواريخ الأحداث: | Date Created: 20240619 Date Completed: 20240619 Latest Revision: 20240622 |
| رمز التحديث: | 20240622 |
| مُعرف محوري في PubMed: | PMC11186949 |
| DOI: | 10.1007/s00253-024-13193-4 |
| PMID: | 38896301 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 1432-0614 |
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| DOI: | 10.1007/s00253-024-13193-4 |