Detection of Adventitious Viruses

A major concern when using mammalian cell lines for production of a biotechnology product is the risk of viral contamination. Such contamination could have serious clinical consequences; in particular, the administration of the contaminated drug could be detrimental to the patient. Drug contamination can arise through the introduction of adventitious (accidentally introduced) viruses during the manufacturing process. The likely sources of contamination include the use of contaminated cell culture media, a breakdown in GMP allowing operator or other external contamination, or the use of contaminated reagents used in the process, such as a monoclonal antibody affinity chromatography column. These contaminations require the development of suitable analytical techniques to ensure the absence of human and animal adventitious viruses.

In Procelltech we perform two techniques:

• in vitro testing (IVVT) by cytopathic effects and/or adsorption of red blood cells on detector cells inoculated with test article and/or supernatants. This is a qualitative assay with a duration of 14 days (EP 6.0., 2.6.7.) or 28 days (USP). This assay is also in compliance to the international guide line ICH Q5A/Q5B and the Center for Biologics Evaluation and Research, FDA, Points to Consider in the Characterization of Cell Lines Used To Produce Biologicals (1993)
• q-PCR evaluation of 13 viruses (BL3)

DETECTION OF RETROVIRUSES (F-PERT)

In Procelltech, to determine the cell culture contamination by retroviruses is performed the Enhanced Product Transcriptase Assay (PERT). This assay is conducted in compliance to European Pharmacopoeia (EP 6.0., 2.6.7.). Since reverse transcriptase is an exclusive enzyme of retroviruses is a specific but indirect indicator of contamination. In Procelltech is performed the quantitative method called fluorescence-PERT (F-PERT). This method uses a standard curve of highly purified reverse transcriptase.

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