Gamma irradiating chromatography columns enables bioburden‐free integrated continuous biomanufacturing

Abstract An important consideration for integrated continuous biomanufacturing is that the downstream chromatography steps integrated with the bioreactor should maintain a low bioburden state throughout the entire duration of the operation. One potential strategy to achieve this is to start bioburden‐free and functionally close the chromatography system. While chromatography skids themselves can be rendered bioburden‐free, limitations exist in applying these methods to chromatography columns. The small column sizes used in continuous multicolumn chromatography enable gamma irradiation of disposable columns to render them bioburden‐free. However, this approach has not been widely implemented, likely because gamma irradiation can negatively impact resin performance. Here, several protective mobile‐phase modifiers were screened and shown to help chromatography resins retain naïve‐like performance. Gamma irradiated columns were then integrated into perfusion bioreactors for continuous capture. Successful integrated continuous capture downstream of perfusion bioreactors for greater than 40 days using protein A, custom affinity, and non‐affinity capture resins for multiple biologic modalities is demonstrated in development and commercial settings. No indications of time‐based performance decline or bioburden growth have been observed. This strategy enables bioburden‐free integrated continuous biomanufacturing operations and could allow full process closure and decreased environmental control requirements for facilities; thus, permitting simultaneous multi‐product operations in a ballroom arrangement.