earticle

영어

In biopharmaceutical manufacturing, many products are produced by Chinese hamster ovary (CHO) cells. Validation of viral safety is essential in ensuring the safety of CHO cell culture-derived biopharmaceuticals, because CHO cells contain endogenous retrovirus like particles(RVLP). RVLP should be removed and/or inactivated during the manufacture of biopharmaceuticals, and must not be present in the final product. Therefore validated quantitative detection method is required for ensuring the safety of biopharmaceuticals against RVLP. In this study, a TaqManbased real-time quantitative PCR assay to detect RVLP was developed and validated. Specific primers for amplification of RVLP nucleic acid was selected, and RVLP RNA was quantified by use of TaqMan probe. The sensitivity of the assay was calculated to be 2.24×100 copies/μL. The TaqMan-based real-time PCR assay was proven to be reproducible and very specific to RVLP. The established real-time PCR assay was successfully applied to the detection of RVLP during the manufacture of a therapeutic antibody using CHO cells. Therefore, it was concluded that this fast, specific, sensitive, and robust assay is invaluable for detection of RVLP and virus clearance evaluation during the manufacture of CHO cell culture-derived biopharmaceuticals.