Investigating Virus Clearance via pH Inactivation During Biomanufacturing

Author

Wenbo XuFollow

Date of Graduation

5-2019

Document Type

Thesis

Degree Name

Bachelor of Science in Biomedical Engineering

Degree Level

Undergraduate

Department

Biomedical Engineering

Advisor/Mentor

Qian, Xianghong

Abstract

In the processing of biopharmaceuticals, viral clearance and viral safety are important for the development of monoclonal antibodies. Murine xenotropic leukemia virus (XMuLV) is one of the retroviruses, recommended by Food and Drug Administration (FDA) as a model virus for viral clearance via inactivation from therapeutics derived from Chinese hamster ovary cells (CHO). A robust and effective method was investigated to clear or inactivate endogenous viruses by low pH inactivation. The effects of different conductivity and inactivated time on XMuLV clearance was determined. Acetate buffer was prepared with different conductivity, and 2% XMuLV was spiked into acetate buffer. XMuLV virus particles could be effectively inactivated in acetate buffer at pH 3.6. According to TICD₅₀ assay, the inactivation time of around 60 minutes was enough to clear all the viruses with more than 4 logs reduction value (LRV). Also, 50 mM acetate buffer has the most rapid inactivation process. TICD₅₀ assays were able to determine the XMuLV virus titer within 95% confidence level, by using 8 replicates and 10-fold series dilution factor.

Keywords

Virus Clearance, XMuLV virus, pH inactivation, Log Reduction Value, PG-4 cell line, Virus titer, service learning

Citation

Xu, W. (2019). Investigating Virus Clearance via pH Inactivation During Biomanufacturing. Biomedical Engineering Undergraduate Honors Theses Retrieved from https://scholarworks.uark.edu/bmeguht/69