Improved method for cloning of antibody secreting cells

抗体分泌细胞克隆的改进方法

• 批准号: 7155325
• 负责人: Gary R. Bright
• 金额: $ 20.64万
• 依托单位: CYNTELLECT, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7155325
• 关键词: antibody; biotherapeutic agent; cell line; clone cells; lasers; proteins; secretion

DESCRIPTION (provided by applicant): Due to the rapidly increasing number of therapeutic humanized monoclonal antibodies being developed and FDA-approved, there is a clear need for improved cell line development to support recombinant therapeutic protein manufacturing. There is a current and projected shortage of worldwide GMP manufacturing capacity for these therapeutic proteins, and approaches for getting more protein from the available bioreactor capacity are deemed critical. Generation of stable high-secreting producer cell lines is a major bottleneck, requiring a 4-10 month process with no guarantee of success. Cloning of cells for high expression of a secreted product (e.g., antibody) is a particular challenge because the secreted product does not remain associated with the cell. The novel laser-enabled analysis and processing (LEAP(tm)) technology platform for high-throughput cell imaging and in situ laser-mediated cell manipulation has the potential to address these needs. Preliminary results shown within indicate significantly improved specific antibody secretion rates from LEAP-derived clones, along with an acceleration of the process, as compared to current methods. The goal of Phase I is to confirm these preliminary results and develop improvements by pursuing the following specific aims: (1) optimize in situ detection of secreted antibody and laser-mediated cell purification; (2) develop automation of cloning processes and monitoring of clone outgrowth; and (3) demonstrate impact of LEAP on cell cloning for recombinant humanized antibody production. Phase I success will lead to a Phase II program to improve, apply and extend the technology, leading to Phase III commercialization with broad impact on recombinant antibody production. In biopharmaceutical manufacturing, over $30 billion worth of therapeutic proteins are produced annually from large-scale culture of recombinant cell lines producing a secreted protein. The number of biopharmaceuticals in clinical development and being FDA approved each year is growing dramatically, resulting in a worldwide shortage of GMP manufacturing capacity for these proteins. Therefore, methods to improve the yield of protein within the current bioreactor capacity constraints are highly desirable.

描述（由申请人提供）：由于正在开发和FDA批准的人性化的单克隆抗体数量迅速增加，因此显然需要改善细胞系发育以支持重组治疗性蛋白质生产。这些治疗蛋白目前存在全球GMP制造能力的短缺，并且认为从可用的生物反应器能力中获取更多蛋白质的方法被认为至关重要。产生稳定的高分子生产商细胞系是一种主要的瓶颈，需要4-10个月的过程，不能保证成功。细胞的克隆用于高表达分泌产物（例如抗体）是一个特殊的挑战，因为分泌的产物与细胞没有关联。用于高通量细胞成像和原位激光介导的细胞操作的新型激光分析和处理（LEAP（TM））技术平台有可能满足这些需求。与当前方法相比，在飞跃衍生的克隆中所显示的初步结果表明，飞跃克隆的特定抗体分泌速率显着提高了特定的抗体分泌速率以及该过程的加速度。第一阶段的目的是通过追求以下特定目的来确认这些初步结果并发展改进：（1）优化原位检测分泌的抗体和激光介导的细胞纯化； （2）发展克隆过程的自动化和克隆生长的监测； （3）显示了LEAP对重组人源化抗体产生的影响。第一阶段的成功将导致II期计划，以改进，应用和扩展技术，从而导致III期商业化，并对重组抗体的产生产生广泛的影响。在生物制药制造中，每年从产生分泌蛋白质的重组细胞系的大规模培养物中生产超过300亿美元的治疗蛋白。每年临床开发和FDA批准的生物药物数量正在急剧增长，从而导致这些蛋白质的GMP制造能力短缺。因此，高度期望在当前生物反应器能力限制内提高蛋白质产量的方法。