SBIR Phase I: Novel Compact Cell Settler for Perfusion Cultures of Microbial Cells

SBIR 第一阶段：用于微生物细胞灌注培养的新型紧凑型细胞沉降器

• 批准号: 1519654
• 负责人: Dhinakar Kompala
• 金额: $ 15万
• 依托单位: Sudhin Biopharma Company
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1519654&HistoricalAwards=false
• 关键词: SBIR; Phase; Novel; Compact; Cell

The broader impact/commercial potential of this Small Business Innovative Research (SBIR) project is to reduce the manufacturing cost of therapeutic biologics using more efficient production technologies. This will be accomplished by developing novel cell retention technologies for increasing cell concentration and productivity in continuous perfusion bioreactor cultures of recombinant microbial and mammalian cells. Based on the inclined settler technology already proven in large scale mammalian cell cultures, compact and more efficiently scalable cell or particle settlers are being developed to expand its applications to other industrial areas, such as microbial yeast secretory production systems, heterogeneous catalysis, municipal water clarification, wastewater treatment, etc. Following thorough characterization of a novel compact cell settler device for recombinant microbial yeast cells during this Phase I grant, it will be launched commercially for large-scale manufacturing of biologics from recombinant microbial and mammalian cells. As biological manufacturing technology matures from early fed-batch cultures to continuous manufacturing due to increasing market size and more consistent product quality-by-design considerations, these novel compact settlers will enable more efficient production of many therapeutic biologics in large scale high cell density perfusion bioreactors and contribute to a significant reduction in cost of their manufacturing from recombinant mammalian and microbial cell cultures.This SBIR Phase I project proposes to develop a novel method of scaling up the proven inclined settler technology, which has been scaled up rectilinearly in the past as lamellar settlers. While the lamellar settlers have been used successfully to manufacture therapeutic biologics secreted by recombinant mammalian cells in high cell density perfusion bioreactors, they have not yet been adapted for microbial cell cultures. The research objectives of this proposal are to develop more compact and efficient settlers for recombinant yeast cells and characterize the capability of these settlers to remove dead cells selectively in the harvest and recycle the live and productive cells back to the bioreactors. Initial results from the first prototype demonstrate selective removal of the smaller yeast cells, while most of the larger cells are concentrated and recycled back to the bioreactor. Lower cell concentrations and smaller size distributions of the yeast cells leaving in the harvest stream from these compact settlers compared to the those in bioreactors will be measured in steady state continuous cultures and dramatically increasing cell density in the perfusion bioreactor due to the complete recycle of live and productive cells to bioreactor will be characterized for different perfusion harvest rates.

该小型企业创新研究（SBIR）项目的更广泛影响/商业潜力是使用更高效的生产技术降低治疗性生物制剂的生产成本。这将通过开发新的细胞保留技术来实现，以增加重组微生物和哺乳动物细胞的连续灌注生物反应器培养中的细胞浓度和生产率。 基于已经在大规模哺乳动物细胞培养中证明的倾斜沉降器技术，正在开发紧凑且更有效地可扩展的细胞或颗粒沉降器以将其应用扩展到其他工业领域，例如微生物酵母分泌生产系统、多相催化、市政水澄清、废水处理、在第一阶段资助期间对用于重组微生物酵母细胞的新型紧凑型细胞沉降器装置进行彻底表征之后，它将投入商业使用，用于从重组微生物和哺乳动物细胞大规模生产生物制剂。 随着生物制造技术从早期的补料分批培养到连续制造的成熟，由于市场规模的增加和更一致的产品质量设计考虑，这些新型紧凑沉降器将使得能够在大规模高细胞密度灌注生物反应器中更有效地生产许多治疗性生物制剂，并有助于显著降低由重组哺乳动物和微生物细胞培养物制造它们的成本。SBIR第一阶段项目提出开发一种扩大经验证的倾斜沉降器技术的新方法，该技术在过去已被直线扩大为层状沉降器。 虽然层状沉降器已成功用于在高细胞密度灌注生物反应器中生产重组哺乳动物细胞分泌的治疗性生物制剂，但它们尚未适应微生物细胞培养。该提案的研究目标是开发更紧凑和有效的重组酵母细胞的沉降器，并表征这些沉降器在收获中选择性地去除死细胞并将活细胞和生产细胞再循环回生物反应器的能力。第一个原型的初步结果表明，较小的酵母细胞被选择性地去除，而大多数较大的细胞被浓缩并回收回生物反应器。 与生物反应器中的那些相比，将在稳态连续培养物中测量从这些紧凑沉降器的收获流中留下的酵母细胞的较低细胞浓度和较小尺寸分布，并且将针对不同的灌注收获速率表征由于活细胞和生产细胞完全再循环至生物反应器而导致的灌注生物反应器中细胞密度的显著增加。