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.

中文翻译：

---

γ辐照色谱柱可实现无生物负荷的集成连续生物制造

集成连续生物制造的重要考虑因素是，与生物反应器集成在一起的下游色谱步骤应在整个操作过程中保持较低的生物负荷状态。实现这一目标的一种潜在策略是启动无生物负荷的功能并关闭色谱系统。尽管色谱撬片本身可以做到无生物负荷，但将这些方法应用于色谱柱仍然存在局限性。连续多柱色谱中使用的较小色谱柱尺寸可对一次性色谱柱进行伽马射线辐照，从而使其无生物负担。但是，该方法尚未得到广泛实施，这可能是因为伽马射线会对树脂性能产生负面影响。这里，筛选了几种保护性流动相改性剂，并证明它们可以帮助色谱树脂保持幼稚的性能。然后将伽马射线辐照的色谱柱整合到灌注生物反应器中以进行连续捕获。在开发和商业环境中，已经证明了使用蛋白A，自定义亲和力和非亲和力捕获树脂对多种生物形态成功进行了灌注生物反应器下游的连续集成捕获超过40天。没有观察到基于时间的性能下降或生物负荷增长的迹象。该策略可实现无生物负荷的集成式连续生物制造操作，并可以完全关闭流程并降低设施的环境控制要求；因此，允许在宴会厅布置中同时进行多产品操作。