Increased production of protein from senescent cells

衰老细胞产生的蛋白质增加

• 批准号: 7053222
• 负责人: Thomas Primiano
• 金额: $ 44.77万
• 依托单位: CDI BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7053222
• 关键词: CHO cells; bioreactors; biotechnology; biotherapeutic agent; cell line; cell senescence; hybridomas; monoclonal antibody; protein biosynthesis; technology /technique development; tissue /cell culture

DESCRIPTION (provided by applicant): Most commercial therapeutic proteins, such as monoclonal antibodies (Mabs), are produced from traditional hybridoma cultures or genetically engineered Chinese hamster ovary (CHO) cells using large scale : bioreactors. Bioprocessing of therapeutic proteins is exceedingly expensive because of the high costs of , building and maintaining FDA-approved facilities, complex serum-free media, and talented labor. There is a tremendous market need for methods that can enhance protein production that ideally complement the current methods of production. The overall goal of the proposed project is to evaluate the effectiveness of senescence-enhanced protein production (RP Shift) in commercially relevant bioreactor setting. In the Phase I portion .of this project, Mab production was enhanced nearly 20-fold from RP Shift-competent hybridoma cells in small flasks. The Phase II portion of this project addresses the effectiveness of the RP Shift in commercially relevant bioreactors, the most commonly used being batch and flow-through systems. The aims of this Phase II proposal are to monitor commercially relevant endpoints from RP Shift-competent CH450 and MH70 hybridoma cells that were engineered in the Phase I portion of the project. Specifically, 1) enhanced productivity of Mab (in pg/cell/day monitored daily), 2) increased culture lifetime measured in days), and 3) the stability and integrity of the Mab produced will be measured. First cell lines with increased productivity of at least 2-fold during RP Shift in small volume test systems will be isolated. Of these RP Shift competent cell lines, the best two will be transferred to commercial scale bioreactors and examined for enhanced protein production and increased bioreactor lifetimes. It is expected that enhancements of at least 2-fold in large-scale bioreactors will attract interest from biopharmaceutical manufacturers. During Phase III, the RP Shift technology and ready-to-go RP Shift-competent cell lines will be licensed to biopharmaceutical manufacturers in an effort to reduce costs for them and ultimately the general public.

描述（由申请人提供）：大多数商业治疗蛋白，例如单克隆抗体（MAB），是由传统的杂交瘤培养物或基因工程性的中国仓鼠卵巢（CHO）细胞生产的：使用大规模：生物反应器。由于建立和维护FDA批准的设施，复杂的无血清媒体和才华横溢的劳动力，因此对治疗蛋白的生物处理非常昂贵。对于可以增强蛋白质生产的方法，理想地可以补充当前生产方法的蛋白质产生。拟议项目的总体目标是评估商业相关的生物反应器设置中衰老增强蛋白（RP转移）的有效性。在该项目的第一阶段部分中，从小烧瓶中的RP换档型杂交瘤细胞增强了MAB的产生近20倍。该项目的第二阶段部分解决了RP转移在商业相关的生物反应器中的有效性，最常用的是批处理和流通系统。该第二阶段提案的目的是监视来自RP转移能力的CH450和MH70杂交瘤细胞的商业相关端点，这些杂交瘤细胞是在项目的I期部分设计的。具体而言，1）MAB的生产率提高（每天监测的PG/Cell/Day中），2）在天数中测量的培养寿命增加），3）将测量产生的MAB的稳定性和完整性。在小体积测试系统中，在RP移位期间，生产率提高至少2倍的第一细胞系将被隔离。在这些RP偏移竞争的细胞系中，最好的两个将被转移到商业尺度生物反应器中，并检查是否有增强的蛋白质产生并增加生物反应器的寿命。预计在大规模生物反应器中至少增强了至少2倍的增强能力，它将吸引生物制药制造商的兴趣。在第三阶段，RP轮班技术和现成的RP转移型细胞系将获得生物制药制造商的许可，以降低他们以及最终的公众成本。