Evaluation of the Virus Elimination and Inactivation at Different Stages of the Model Biotechnological Process for the Production of a Drug Based on the Monoclonal Antibody Fab-Fragment

Abstract

Virus safety insuring is one of the most serious problems in the development of biotechnological drugs produced using animal cell lines. Quantitative assessment of virus removal and inactivation is an integral approach to show the reliability of the target compound production. In this work, the model experiments have been carried out on the purification of the monoclonal antibody recombinant Fab-fragment from viruses of various origins and properties: xenotropic murine leukemia virus (X-MuLV), pseudorabies virus (PRV), Reo-3 virus and encephalomyocarditis virus (EMCV). It was shown that two methods, acidification to pH 3.2 and nanofiltration, made it possible to reduce the virus infectivity to levels undetectable in cell cultures (according to TCID50). The developed multistage purification process of the target protein provided an overall decrease in viral clearance to the following values: X-MuLV≥10.17lg, PRV≥13.98lg, Reo-3≥8.09lg and EMCV≥4.98lg. These results confirm that the developed technology ensures the virus safety during the production of a monoclonal antibody recombinant Fab-fragment by CHO cell line. These results confirm virus safety of production technology of recombinant monoclonal antibody Fab-fragment produced in CHO cell line. virus safety, virus elimination, virus inactivation, nanofiltration, Fab-fragment, monoclonal antibody, CHO cell line