Application of cation exchange chromatography in bind and elute and flowthrough mode for the purification of enteroviruses.

Members of the enterovirus genus are promising oncolytic agents. Their morphogenesis involves the generation of both genome-packed infectious capsids and empty capsids. The latter are typically considered as an impurity in need of removal from the final product. The separation of empty and full capsids can take place with centrifugation methods, which are of low throughput and poorly scalable, or scalable chromatographic processes, which typically require peak cutting and a significant trade-off between purity and yield. Here we demonstrate the application of packed bed cation exchange (CEX) column chromatography for the separation of empty capsids from infectious virions for a prototype strain of Coxsackievirus A21. This separation was developed using high throughput chromatography techniques and scaled up as a bind and elute polishing step. The separation was robust over a wide range of operating conditions and returned highly resolved empty and full capsids. The CEX step could be operated in bind and elute or flowthrough mode with similar selectivity and returned yields greater than 70% for full mature virus particles. Similar performance was also achieved using a selection of other bead based CEX chromatography media, demonstrating general applicability of this type of chromatography for Coxsackievirus A21 purification. These results highlight the wide applicability and excellent performance of CEX chromatography for the purification of enteroviruses, such as Coxsackievirus A21.

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Copyright © 2022. Published by Elsevier B.V.

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Declaration of Competing Interest Spyridon Konstantinidis reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Murphy R Poplyk reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Andrew R Swartz reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Richard R Rustandi reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Rachel Thompson reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Sheng-Ching Wang reports a relationship with Merck & Co Inc that includes: employment and equity or stocks. Spyridon Konstantinidis has patent #US20210187049A1 pending to Merck Sharp and Dohme Corp. Murphy R Poplyk has patent #US20210187049A1 pending to Merck Sharp and Dohme Corp. Andrew R Swartz has patent #US20210187049A1 pending to Merck Sharp and Dohme Corp.