Platform for Rapid Expression, Purification, and Analysis of Patient-Specific Gene Therapy Products

患者特异性基因治疗产品快速表达、纯化和分析的平台

• 批准号: 10414307
• 负责人: Michael Daniele
• 金额: $ 58.17万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414307
• 关键词: Platform; Rapid; Expression; Purification; Analysis

ABSTRCT By 2050, adeno-associated viruses (AAVs) will be established as the workhorse for in vivo gene therapy, impacting 60 - 80 million patients per year worldwide, and the success of this gene therapy will rely on the ability to manufacture application-/patient-specific AAVs affordably and rapidly. However, while in vivo and ex vivo gene therapy products are advancing rapidly, their biomanufacturing technology has not seen comparable progress. At present, manufacturers achieve the mandated product purity upon strenuous optimization - at the expense of productivity and product efficacy and run multiple production batches to meet the clinical demand -with significant delays and cost to patients (up to $1.2M per individual). To address the challenges faced by current AAV biomanufacturing, we propose to develop an integrated purification and sensing technologies to accelerate process design and optimization and make small-batch AAV production affordable to specific patient cohorts. The technology is based on the integration of chromatographic adsorbents for isolation of “full” AAVs in flow- through mode coupled with label-free impedimetric biosensors for quantifying AAV titer and full:empty ratio. These technologies will enable the application of advanced analytics for the prediction of AAV therapeutic efficacy by correlating process parameters with biomolecular-level CQAs of AAVs. The integrated purification and biosensor technology will be demonstrated in a pilot process for producing patient-specific doses of rAAV2- retro and scAAV9-CB-CLN6, which have been shown to correct brain and behavioral pathologies in CLN6- Batten disease, a rare lysosomal storage disease.

中文摘要 到2050年，腺相关病毒（AAV）将成为体内基因治疗的主力， 每年影响全球6000 - 8000万患者，这种基因疗法的成功将取决于 以经济和快速地制造应用/患者特异性AAV。然而，尽管体内和离体基因 治疗产品发展迅速，但其生物制造技术却没有取得可比的进展。 目前，制造商通过艰苦的优化来实现规定的产品纯度-以牺牲 生产效率和产品功效，并运行多个生产批次以满足临床需求-具有显著的 延误和患者成本（每人高达120万美元）。为了应对当前AAV面临的挑战， 生物制造，我们建议开发一个综合的净化和传感技术，以加速 这将有助于进行工艺设计和优化，并使特定患者群体能够负担得起小批量AAV生产。 该技术基于色谱吸附剂的集成，用于在流动中分离“完整”AAV， 通过模式与用于定量AAV滴度和满：空比率的无标记阻抗生物传感器偶联。 这些技术将使先进的分析技术能够应用于预测AAV治疗性疾病。 通过将工艺参数与AAV的生物分子水平的CQA相关联来确定有效性。综合净化 和生物传感器技术将在生产患者特异性剂量的rAAV 2- retro和scAAV 9-CB-CLN 6，它们已被证明能纠正CLN 6-CLN 8中的大脑和行为病理。 Batten病，一种罕见的溶酶体贮积病。