Prediction of viral filtration performance of monoclonal antibodies based on biophysical properties of feed

Abstract

Controlling viral contamination is an important issue in the process development of monoclonal antibodies (MAbs) produced from mammalian cell lines. Virus filtration (VF) has been demonstrated to be a robust and effective clearance step which can provide ≥4 logs of reduction via size exclusion. The minimization of VF area by increasing flux and filter loading is critical to achieving cost targets as VFs are single use and often represent up to 10% of total purification costs. The research presented in this publication describes a development strategy focused on biophysical attributes of product streams that are directly applicable to VF process performance. This article summarizes a case study where biophysical tools (high-pressure size exclusion chromatography, dynamic light scattering, and absolute size exclusion chromatography) were applied to a specific MAb program to illustrate how changes in feed composition (pH, sodium chloride concentration, and buffer salt type) can change biophysical properties which correlate with VF performance. The approach was subsequently refined and expanded over the course of development of three MAbs where performance metrics (i.e., loading and flux) were evaluated for two specific virus filters (Viresolve Pro and Planova 20N) during both unspiked control runs and virus clearance experiments. The analyses of feed attributes can be applied to a decision tree to guide the recommendation of a VF filter and operating conditions for use in future MAb program development. The understanding of the biophysical properties of the feed can be correlated to virus filter performance to significantly reduce the mass of product, time, and costs associated with virus filter step development.

The effects of buffer condition on the fouling behavior of MVM virus filtration of an Fc‐fusion protein

“…Previous studies (Bakhshayeshi & Zydney, 2008; G. R. Bolton, Spector, & LaCasse, 2006;Dishari et al, 2015;Hongo-Hirasaki, Komuro, & Ide, 2010;Rayfield et al, 2015;Wickramasinghe, Stump, Grzenia, Husson, & Pellegrino, 2010) show that buffer conditions, including pH, buffer type, and ionic strength could affect the filtration performance of the virus filters. In addition, earlier studies have shown that the performance of virus filters is also dependent on the specific protein in the feed (Stuckey et al, 2014).…”

“…Many previous studies have investigated the effect of pH and types of fouling species on the virus filter capacity and proposed various strategies such as using adsorptive prefilters to achieve higher throughput (Bakhshayeshi & Zydney, 2008; G. R. Dishari et al, 2015;Hongo-Hirasaki et al, 2010;Wickramasinghe et al, 2010). Previous work (Rayfield et al, 2015) also examined the correlation between the biophysical properties and the virus filtration performance and found that molecular size determined by absolute size-exclusion chromatography is an important factor. Previous work (Rayfield et al, 2015) also examined the correlation between the biophysical properties and the virus filtration performance and found that molecular size determined by absolute size-exclusion chromatography is an important factor.…”

Use of a noninfectious surrogate to predict minute virus of mice removal during nanofiltration

“…Bacteriophages, endogenous CHO Type C retrovirus‐like particles (RVLPs), and biologic feedstreams have been used in attempts to economically predict the outcomes of viral clearance studies . However, due to the differences in physicochemical characteristics of bacteriophage models and the complexities involved in producing purified and highly concentrated RVLP spiking stocks, the scope of utility for these surrogates have remained narrow.…”

Suitability of a generic virus safety evaluation for monoclonal antibody investigational new drug applications

“…It has been shown that virus breakthrough can occur with small virus retentive filters during long processing times . However, proper evaluation and control of process parameters and solution properties can maintain high filter flux and mitigate this risk . These findings suggest that when an operating space is determined, within which there is sufficient clearance of small virus to assure patient safety, it may be generically applied to larger virus across multiple products.…”