Chromatographic removal combined with heat, acid and chaotropic inactivation of four model viruses

Abstract

The virus removal of protein A affinity chromatography, inactivation capacity, acid pH and a combination of high temperature with a chaotropic agent was determined in this work. The model viruses studied were sendaivirus, human immunodeficency virus (HIV-IIIb), human poliovirus type-II, human herpesvirus I and canine parvovirus. The protein A affinity chromatography showed a maximum reduction factor of 8 logs in the case of viruses larger than 120 nm size, while for small viruses (18-30 nm) the maximum reduction factor was about 5 logs. Non viral inactivation was observed during the monoclonal antibody elution step. Low pH treatment showed a maximum inactivation factor of 7.1 logs for enveloped viruses. However, a weak inactivation factor (3.4 logs) was obtained for DNA nonenveloped viruses. The combination of high temperature with 3 M KSCN showed a high inactivation factor for all of the viruses studied. The total clearance factor was 23.1, 15.1, 13.6, 20.0 and 16.0 logs for sendaivirus, HIV-IIIb, human poliovirus type-II, human herpesvirus I and canine parvovirus, respectively.

A Novel, Q‐PCR Based Approach to Measuring Endogenous Retroviral Clearance by Capture Protein A Chromatography

“…The clearance results obtained using both assays, in both the process characterization and resin reuse studies were similar. Overall, the combined data provide confirmation that protein A is a robust virus removal step (Brorson et al, 2003a;Valdes et al, 2002).…”

Factorial screening of antibody purification processes using three chromatography steps without protein A

“…For example, the ability of the S-Q-HIC, S-Q-ABx, and S-ABx-Q processes to remove host cell DNA, small molecules, and antibody variants must be evaluated, and process robustness and process economics must be investigated. Many of these chromatography steps may not provide as much virus clearance as protein A but several alternative methods, such as filtration or chemical inactivation, may be employed [18,19,31,32]. Although most of the resins are stable to sodium hydroxide, resin cleanability must be investigated.…”

“…Affinity chromatography also provides significant virus clearance [18,19]. Because it binds a variety of mammalian IgG molecules [20], protein A affinity chromatography also allows process harmonization for multi-product manufacturing.…”

“Quality Tools for Improving a System of documentation as the basis for Good Manufacturing Practices”. Practical Case

“…After this chromatography, incubation in 0.1 molar (M) citric acid pH 3.0 for an hour at 4°C was carried out. The ultra filtration and sterile filtration steps complete the basic diagram of the purification process [4].…”