Evaluation of viral clearance in the production of HPV-16 L1 virus-like particles purified from insect cell cultures
| Autor: | Sue-Nie Park, Jei-Jun Bae, Young-Lim Kim, Byoung-Guk Kim, Hong Ki Min, Soon-Nam Kim, Hye-Sung Jeong, Jung-Yun Choi, Seung-Rel Ryu, Hong-Jin Kim, Jin-Ho Shin |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Insecta
Lysis viruses Cell Culture Techniques Bioengineering Applied Microbiology and Biotechnology Virus law.invention Microbiology Viral envelope law Animals Centrifugation Pharmacology Infectivity Human papillomavirus 16 General Immunology and Microbiology biology General Medicine biology.organism_classification Virology Cell culture DNA Viral Recombinant DNA RNA Viral Minute virus of mice Biotechnology |
| Zdroj: | Biologicals. 34:273-279 |
| ISSN: | 1045-1056 |
| DOI: | 10.1016/j.biologicals.2005.11.008 |
| Popis: | Biopharmaceutical products produced from cell cultures have a potential for viral contamination from cell sources or from adventitious introduction during production. The objective of this study was to assess viral clearance in the production of insect cell-derived recombinant human papillomavirus (HPV)-16 type L1 virus-like particles (VLPs). We selected Japanese encephalitis virus (JEV), bovine viral diarrhea virus (BVDV), and minute virus of mice (MVM) as relevant viruses to achieve the aim of this study. A downstream process for the production of purified HPV-16 L1 VLPs consisted of detergent lysis of harvested cells, sonication, sucrose cushion centrifugation, and cesium chloride (CsCl) equilibrium density centrifugation. The capacity of each purification/treatment step to clear viruses was expressed as reduction factor by measuring the difference in log virus infectivity of sample pools before and after each process. As a result, detergent treatment (0.5% v/v, Nonidet P-40/phosphate-buffered saline) was effective for inactivating enveloped viruses such as JEV and BVDV, but no significant reduction (1.0 log(10)) was observed in the non-enveloped MVM. The CsCl equilibrium density centrifugation was fairly effective for separating all three relevant adventitious viruses with different CsCl buoyant density from that of HPV-16 L1 VLPs (JEV, BVDV, and MVM = 4.30, 3.10,or = 4.40 log(10) reductions). Given the study conditions we used, overall cumulative reduction factors for clearance of JEV, BVDV, and MVM wereor = 10.50,or = 9.20, andor = 6.40 log(10) in 150 ml of starting cell cultures, respectively. |
| Databáze: | OpenAIRE |
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