SBIR Phase I: Novel compact cell settlers for high cell density perfusion cultures of mammalian cells

该小型企业创新研究（SBIR）项目的更广泛影响/商业潜力是开发一种新型紧凑型细胞沉降器，该沉降器可以选择性地从细胞培养液中去除死细胞和细胞碎片，并将所有活细胞和生产细胞完全回收回连续灌注生物反应器。 由于这种无膜装置不像竞争的基于膜的细胞保留装置那样易于堵塞，因此不需要在3或4周后过早终止单个灌注生物反应器培养（由于死细胞和细胞碎片的积累导致产品质量降低和当前装置中的膜堵塞）。 与其他细胞保留装置相比，新型紧凑型细胞沉降器具有较小的尺寸和占地面积，并且增加了大量生物制造公司更广泛地适应该技术的可能性，并降低了治疗性生物制剂的制造成本。 由于生物类似药的价格竞争预计将成功的产品，即将摆脱专利保护，降低其生产成本，使用上级细胞培养技术将使这些药物的负担能力增加到更广泛的患者群体。该SBIR第一阶段项目提出开发一种新颖的紧凑且更有效的方法来扩大倾斜沉降器技术，该技术已被低效地扩大为直线层状沉降器。 虽然一些生物制造商已经成功地使突出的层状沉降器适应于在高细胞密度灌注生物反应器中由重组哺乳动物细胞制造治疗性生物制剂，但是预测所提出的新型紧凑型细胞沉降器以与原始倾斜层状沉降器设计相比小得多的占地面积实现从高细胞密度灌注生物反应器中分离死细胞和细胞碎片的所需分离。该研究的三个关键目标是：1）证明新型紧凑型细胞沉降器可以无限期地实现和保持高活细胞密度，2）确定两种不同尺寸的较小沉降器无限期地保持高活细胞密度的最大灌注速率，以及3）开发新的缩放定律，可以预测较大商业规模生物反应器的沉降器的尺寸要求。 这些目标将通过系统的实验计划来实现，以研究两种不同尺寸和两种不同哺乳动物细胞类型的新型沉降器的性能。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is the development of a novel compact cell settler that can selectively remove dead cells and cell debris from the cell culture broth and completely recycle all the live and productive cells back to the continuous perfusion bioreactor. As this membrane-free device is not prone to clogging like the competing membrane-based cell retention devices, a single perfusion bioreactor culture does not need to be terminated prematurely after 3 or 4 weeks (due to accumulation of dead cells and cell debris resulting in reduced product quality and clogged membranes in current devices). The novel compact cell settler has a small size and footprint compared with other cell retention devices, and increases the likelihood of wider adaptation of the technology by a larger number of biomanufacturing companies and reduces the cost of manufacture of therapeutic biologics. As biosimilar price competition is expected for successful products that are coming off patent protection, reducing the cost of their manufacture using superior cell culture techniques will enable increased affordability of these drugs to a wider patient population. This SBIR Phase I project proposes to develop a novel compact and more efficient method of scaling up inclined settler technology, which has been scaled up inefficiently as rectilinear lamellar settlers. While a few biological manufacturers have successfully adapted the protruding lamellar settlers to manufacture therapeutic biologics from recombinant mammalian cells in high cell density perfusion bioreactors, the proposed novel compact cell settler is predicted to achieve the required separation of dead cells and cell debris from a high cell density perfusion bioreactor with a much smaller footprint compared to the original inclined lamellar settler design. Three key objectives for the research are to 1) demonstrate that the novel compact cell settler can achieve and maintain high viable cell densities indefinitely, 2) determine the maximum perfusion rate that maintain the high viable cell density indefinitely for smaller settlers of two different sizes, and 3) develop new scaling laws that can predict the size requirements of the settlers for larger commercial-scale bioreactors. These objectives will be accomplished by a systematic experimental plan to investigate the performance of the novel settlers at two different sizes and for two different mammalian cell types.