title: Evaluation of viral clearance in the production of HPV-16 L1 virus-like particles purified from insect cell cultures.
abstract: 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 were > or = 10.50, > or = 9.20, and > or = 6.40 log(10) in 150 ml of starting cell cultures, respectively.
authors.affiliation: College of Pharmacy, Chung-Ang University, #221 Huksuk-Dong, Dongjak-Ku, Seoul 156-756, Republic of Korea.
authors.forename: Hye-Sung
authors.initials: HS
authors.lastname: Jeong
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Shin
authors.forename: Jung-Yun
authors.initials: JY
authors.lastname: Choi
authors.forename: Young-Lim
authors.initials: YL
authors.lastname: Kim
authors.forename: Jei-Jun
authors.initials: JJ
authors.lastname: Bae
authors.forename: Byoung-Guk
authors.initials: BG
authors.lastname: Kim
authors.forename: Seung-Rel
authors.initials: SR
authors.lastname: Ryu
authors.forename: Soon-Nam
authors.initials: SN
authors.lastname: Kim
authors.forename: Hong-Ki
authors.initials: HK
authors.lastname: Min
authors.forename: Hong-Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Sue-Nie
authors.initials: SN
authors.lastname: Park
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004279:DNA, Viral / Q000302:isolation & purification; D052162:Human papillomavirus 16 / Q000254:growth & development / Q000302:isolation & purification*; D007313:Insecta / Q000821:virology*; D012367:RNA, Viral / Q000302:isolation & purification
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral