Clearance of porcine circovirus and porcine parvovirus from porcine‐derived pepsin by low pH inactivation and cation exchange chromatography

• Published in: Biotechnology progress 2020-07, Vol.36 (4), p.e2968-n/a
• Main Authors: Yang, Bin, Wang, Hua, Kaleas, Kimberly, Butler, Michelle, Franklin, Jayme, Bill, Amber, Baylis, Sally A., Chen, Qi, Blümel, Johannes
• Format: Article
• Language: English
• Subjects: anion exchange (AEX) chromatography; Cation exchange; cation exchange (CEX) chromatography; Cation exchanging; Chromatography; Contamination; Deactivation; Inactivation; Intermediates; minute virus of mice (MVM); Parvoviruses; Pepsin; pH effects; porcine circovirus (PCV); porcine circovirus Type 1 (PCV1); porcine circovirus Type 2 (PCV2); porcine circovirus Type 3 (PCV3); Pore size; Porosity; Process parameters; Production methods; Resins; Rotavirus; Safety barriers; Trypsin; Vaccines; viral clearance; virus chromatography; Viruses
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1002/btpr.2968

The contamination of oral rotavirus vaccines by porcine circovirus (PCV) raised questions about potential PCV contamination of other biological products when porcine trypsin or pepsin is used in production process. Several methods can be potentially implemented as a safety barrier when animal derived trypsin or pepsin is used. Removal of PCV is difficult by the commonly used viral filters with the pore size cutoff of approximately 20 nm because of the smaller size of PCV particles that are around 17 nm. It was speculated that operating the chromatography step at a pH higher than pepsin's low pI, but lower than pIs, of most viruses would allow the pepsin to flow through the resin and be recovered from the flow through pool whilst the viruses would be retained on the resin. In this study, we investigated low pH inactivation of viruses including PCV Type 1 (PCV1) and PCV1 removal by cation exchange chromatography (CEX) in the presence of pepsin. Both parvovirus and PCV1 could be effectively inactivated by low pH and PCV1 could be removed by POROS 50HS CEX. The POROS 50HS method presented in this article is helpful for designing other CEX methods for the same purpose and not much difference would be expected for similar product intermediates and same process parameters. While the effectiveness needs to be confirmed for specific applications, the results demonstrate that both low pH (pH 1.7) and CEX methods were successful in eliminating PCV1 and thus either can be considered as an effective virus barrier.