Evaluation of viral clearance in the production of HPV-16 L1 virus-like particles purified from insect cell cultures.
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.
Jeong
Hye-Sung
HS
College of Pharmacy, Chung-Ang University, #221 Huksuk-Dong, Dongjak-Ku, Seoul 156-756, Republic of Korea.
Shin
Jin-Ho
JH
Choi
Jung-Yun
JY
Kim
Young-Lim
YL
Bae
Jei-Jun
JJ
Kim
Byoung-Guk
BG
Ryu
Seung-Rel
SR
Kim
Soon-Nam
SN
Min
Hong-Ki
HK
Kim
Hong-Jin
HJ
Park
Sue-Nie
SN
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
D004279:DNA, Viral; D012367:RNA, Viral