Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 15 5 2023 10 12 711 722 EN Seyed Mohammad Rasouli-Nejad Mousavi Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences & Biotechnology, University of Shahid Beheshti, Tehran, Iran Seyed Masoud Hosseini Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences & Biotechnology, University of Shahid Beheshti, Tehran, Iran Samira Ansari CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran 2023 05 30 2023 10 12 Background and Objectives: Viral clearance studies are an essential part of a manufacturer's plan to ensure the safety of an injectable biologic product. In this way, viral safety is a critical quality attribute for biologics such as monoclonal antibodies (Mabs). Evaluation of virus purification by downstream processes is a key component of risk mitigation. In this study, the capability of continuous monoclonal antibody purification steps was evaluated in the process of instant monoclonal antibody purification in different stages of purification, and the amount of reduction or inactivation of each step was determined. Materials and Methods: Four enveloped and non-enveloped viral models VSV, Reovirus, EMCV, and HSV1 were used for spiking in selected samples in the designated tests, to have a comprehensive examination of the ability to clear the virus such as the type of genetic material, chemical resistance, and particle size. A TCID50 and qPCR methods were used to measure viral reduction. Two cell lines, Vero (African green monkey kidney) and L929 (Mouse fibroblast) were used for 4 model viruses propagation. The steps that were evaluated included 4 steps monoclonal antibody purification; cation exchange chromatography, acidic pH treatment, affinity chromatography, and nanofiltration. Results: The nano-filter stage showed the highest viral reduction and cation exchange chromatography showed the lowest reduction. The cumulative decrease using TCID50 is equal to 19.27 [log10] for all steps and for the qPCR method is equal to 12.47 [log10] in three steps of nano-filter, affinity chromatography, and ion exchange chromatography. Conclusion: The overall average reduction coefficient for all four model viruses is significantly high, which indicates the high capacity of the monoclonal antibody production process in inactivating and removing viruses leads to reducing the load of all four model viruses. https://ijm.tums.ac.ir/index.php/ijm/article/view/4222 https://ijm.tums.ac.ir/index.php/ijm/article/download/4222/1617