Reducing a major cost burden of therapeutic cell manufacturing by selectivelyremoving toxic culture byproducts to allow recycling of media

摘要： 基于细胞的疗法制造的巨大成本是成功的重大威胁。 细胞/基因治疗。单独的细胞培养基可以贡献约30-40%的生产成本。 货物（COG）。这是因为，目前的细胞培养方法需要频繁和完全的培养基 替换以从培养物中去除有毒代谢物。这种完全取代媒介的做法 使用新鲜培养基会增加显著的费用并造成不必要的浪费。恢复营养，渗透， 并且pH平衡不需要更换整个培养基体积。在生产规模上，一些 版本的介质将需要回收，以减少浪费和制造的总成本。的目的 目前的研究是开发一种具有成本效益的介质过滤系统，使用椰子壳衍生的活性炭 （CS-AC）可以选择性去除其中两种重要的有毒代谢产物，例如乳酸和铵 细胞培养基。利用这种过滤系统，可以选择性地去除代谢废物，而不污染环境。 代谢成分如缓冲剂和生长因子可以保留，而耗尽的营养物质如糖 并且氨基酸可以在它们被细胞消耗时被补充。确定回收水平， 通过这种过滤技术，对于培养基的每种组分， 复杂的分析由废介质分析的经验数据提供信息。 本研究的最终目的是利用这种过滤技术，开发出一种商业级的 通过将AC嵌入纤维素基质或中性级树脂中，实现一次性过滤装置。的应用 从废培养基中去除有毒代谢物的新过滤装置将改变目前的 正在研究在基于细胞的治疗制造期间减少COG的方法。这种过滤 该系统为负担得起的细胞/基因疗法治疗可能性提供了巨大的希望，预计将具有 对临床医学产生巨大的经济影响。

ABSTRACT: The tremendous cost of cell-based therapies manufacturing is the significant threat to the success of cell/gene therapy treatments. The cell culture medium alone can contribute to around 30-40% of the Cost of Goods (COGs). This is because, the current cell culture methods require frequent and complete media replacement in order to remove toxic metabolites from culture. This practice of completely replacing the medium with fresh medium adds significant expense and contributes to unnecessary waste. Restoring nutrient, osmotic, and pH balance need not require replacement of the entire culture medium volume. At production scale, some version of medium recycling will be required to reduce waste and overall cost of manufacturing. The aim of the current study is to develop a cost-effective media filtration system using coconut-shell derived activated carbons (CS-AC) that can selectively remove two important toxic metabolites such as lactic acid and ammonium from cell culture medium. With such filtration system, metabolic waste products can selectively be removed, non- metabolized components like buffers, and growth factors can be retained, and the depleted nutrients like sugars and amino acids can be replenished as they are consumed by the cells. Determining the level of recycling that is achievable by this filtration technology for each component of the culture medium requires a significantly more complex analysis informed by empirical data from spent medium analysis. The ultimate goal of this study is to utilize this filtration technology and develop a commercial grade single-use filtration device by embedding the AC into cellulose matrix or neutral-grade resins. The application of the new filtration device to remove toxic metabolites from spent culture medium will transform the current approaches being investigated for COG reduction during cell based therapeutic manufacturing. Such a filtration system holds a great promise for affordable cell/gene therapy treatment possibilities, and is expected to have a strong economical impact on clinical medicine.