RESUMEN
Virus filtration has been demonstrated to be an effective and robust dedicated viral clearance step that is used in biopharmaceutical manufacturing processes. Here we present virus filtration data from a multicompany collaboration with data compiled from WuXi Advanced Therapies' and Charles River Laboratories' internal viral clearance databases spanning more than 25 years. The data were sorted by virus removal and type and then further subdivided into murine leukemia virus only, pseudorabies virus only, and reovirus type 3 only categories to allow for analyses of viral clearance results. A total of 2311 virus filtrations were analyzed, composed of 1516 murine leukemia virus, 385 pseudorabies virus, and 410 reovirus type 3 virus filtrations. These data provide clear evidence that will help supplement both internal and industry-wide initiatives focused on using prior knowledge for the creation of modular claims for small virus retentive filters and allow better allocations of resources typically spent on potentially unnecessary studies.
Asunto(s)
Productos Biológicos , Virus , Animales , Virus ADN , Filtración/métodos , Virus de la Leucemia Murina , RatonesRESUMEN
Virus retention filtration is a common step in modern biopharmaceutical manufacturing as it enables efficient removal of potential adventitious and endogenous viruses via size exclusion. Modern parvovirus retention filters have significantly improved fluxes and parvovirus retention in comparison to earlier versions of these filters. However, these filters may be more susceptible to premature fouling and require more effort for process optimization. Here, we demonstrate that polyamide-6,6 (nylon-6,6) membranes when used as prefilters can increase the capacity of these Parvovirus retentive filters that are less susceptible to premature fouling. We found that the mechanism of polyamide-mediated filtration improvement can be explained by the binding of monoclonal antibody (mAb) aggregates with a diameter of 20-100 nm, and we show that this mechanism is shared by other types of adsorptive prefilters. Finally, by the combination of mobile phase screening, additive spiking, and molecular dynamics simulations, we show that polyamide-6,6 removes mAb aggregates through hydrophobic interactions making its design space potentially complementary to other available prefilters. Our studies support the aggregate-mediated mechanism of flux decay during viral filtration and suggest that polyamide-6,6 could be considered as an alternative cost-effective option to extend the capacity of viral filters.
Asunto(s)
Anticuerpos Monoclonales/química , Anticuerpos Antivirales/química , Caprolactama/análogos & derivados , Simulación de Dinámica Molecular , Parvovirus/química , Polímeros/química , Caprolactama/química , FiltraciónRESUMEN
Low pH inactivation of enveloped viruses has historically been shown to be an effective viral inactivation step in biopharmaceutical manufacturing. To date, most statistical analyses supporting modular low pH viral inactivation claims have used descriptive statistical analyses, which in many cases do not allow for probabilistic characterization of future experimental log10 reduction values (LRVs). Using Bayesian hierarchical logistic regression modeling, probability statements regarding the likelihood of successful low pH viral inactivation based on only certain process parameter settings can be derived. This type of analysis also permits statistical modeling in the presence of historical data from different experiments and right-censored data, two issues that have not as yet been satisfactorily dealt with in the literature. The characterization of the probability of successful inactivation allows creation of a modular claim stating future LRVs will be greater than or equal to some critical value, based on only certain process parameter settings of the viral inactivation unit operation. This risk-based approach, when used in conjunction with traditional descriptive statistics, facilitates coherent and cogent decision-making about modular viral clearance LRV claims.LAY ABSTRACT: Viral contamination of biologically derived drug products is a safety concern for both regulatory agencies and drug manufacturers. Validation of the removal and inactivation of model viruses is required to ensure the safety of patients receiving these drugs, and dedicated steps, including viral filtration and chemical inactivation, are often added to manufacturing processes to provide additional clearance and inactivation capabilities. One of these steps, low pH inactivation, exposes enveloped viruses to a low pH environment to reduce the potential of the virus to infect host cells. Because the viral inactivation capability of this well-understood unit operation has been demonstrated for years across many different biological drugs, many companies have begun investigating the use of the modular viral clearance claim for the low pH inactivation step. Modular claims ensure, without experimentation, that a certain level of reduction of virus will occur if specific parameters are used in the manufacturing process, allowing manufacturers to save both time and resources in the early developmental phases of biologically derived drugs. A novel type of statistical analysis is outlined in this article that when used in addition to previously used analyses allows drug manufacturers to estimate a more valid level of virus reduction in modular viral clearance claims.