Rapid pre-release sterility test for PET Drugs.

PET 药物的快速发布前无菌测试。

• 批准号: 10258453
• 负责人: Arkadij Elizarov
• 金额: $ 16.81万
• 依托单位: TRACE-ABILITY, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10258453
• 关键词: Rapid; pre; release; sterility; test

Abstract Traditional process of sterility testing requires a 14-day sample incubation in different media. That makes it impossible to establish a pre-release sterility testing for PET tracers, which have half-lives of 9 to 110 minutes and must be released within 1 hour after synthesis. Moreover, even with numerous controls in place, in the past two years two of three of the largest US PET tracer manufacturers have been issued sterility warnings from the FDA. Trace-Ability proposes a novel approach to rapid sterility testing without incubation, with sterility results available within 1 hour. The solution will take advantage of microfluidic chip technology and will be completely automated and incorporated into the only commercially available and validated automated PET tracer QC platform. Proposed rapid sterility test involves assessing viability by probing whether cellular membranes are intact, and whether there is DNA in those membranes. This will be achieved by selective fluorescent staining of up to 250 µL of sample on a microfluidic chip. To check cell viability, we will take advantage of two stains. The difference of the two signals will indicate whether the organism may be able to reproduce. Proposed solution is intended to enable automated sterility testing and include it into pre-release automated QC workflows for the 1st time in history of PET. Specific Aim 1: Selection of fluorescent stains that can provide the high selectivity and differentiability of viable cells. A combination of stains will be selected that can differentiate between viable and non-viable organisms for 10 typical bacterial species. Evaluation criteria: (1) Viability stains incorporate within 30 minutes, (2) Organisms retain viability after staining, (3) Stains for nonviable organisms have twice the binding affinity of the viability stains, being able to displace any free DNA. (4) bound:unbound ratio > 1000. (5) Limits of Detection down to 10 CFU (colony-forming unit). Specific Aim 2: Development of microfluidic analysis chip for Signal to Noise (S/N) ratio optimization. Taking the optimal stains established in aim 1, these will then be mixed with bacteria in low concentration and loaded onto microfluidic chips with varying channel sizes and shapes while monitoring fluorescence emission for background reduction and S/N increase. Evaluation criteria: (1) a limit of detection (LOD) of 1 CFU, (2) a 100-fold improvement in S/N, (3) filling times less than 30 minutes for 250 μL. Once feasibility is verified in Phase 1, this work has a clear path to commercialization by implementing the test into the Tracer-QC platform. This will allow PET manufactures to have a complete QC package in a single automated system that meets all compliance criteria. This work can extend well beyond PET to compounding pharmacies and other small-scale manufacturers, offering simplification of the experimental setup, procedural changes in production, and regulatory changes to improve patient safety.

抽象的 传统的无菌测试过程需要在不同培养基中进行14天的样品孵育。那 使得不可能为宠物示踪剂建立预释放的不育测试，该测试的半衰期为9至 110分钟，必须在合成后1小时内释放。而且，即使有许多控件 在过去的两年中 FDA的不育警告。 痕量性建议是一种不孵育的新型方法，用于快速无菌测试，不育 结果可在1小时内提供。该解决方案将利用微流体芯片技术，并将 完全自动化并将其纳入唯一的市售和验证的自动化 宠物示踪剂QC平台。拟议的快速无菌测试涉及评估可行性，通过探测是否是否 细胞膜完整，以及这些膜中是否有DNA。这将通过 在微流体芯片上，选择性荧光染色最多250 µl样品。要查看细胞生存能力，我们 将利用两种污渍。两个信号的差异将表明生物体是否可以 能够复制。建议的解决方案旨在实现自动无菌测试，并将其包括在 宠物历史上的第一次释放QC的自动化质量控制工作流程。 特定目的1：选择可以提供高选择性和的荧光污渍 可行细胞的不同性。将选择可以区分可行的污渍的组合 和10种典型细菌的不可生存的生物。评估标准：（1）可行性保留率 在30分钟内掺入（2）染色后的生物保留生存能力，（3）不可行的污渍 生物体具有可行性保留的结合亲和力的两倍，能够取代任何自由DNA。 （4） 绑定：未结合比> 1000。（5）检测限制至10 CFU（菌落形成单元）。具体目标 2：开发信号与噪声（S/N）比率优化的微流体分析芯片。接受 在AIM 1中建立的最佳污渍，然后将它们与低浓度的细菌混合并加载 在监测荧光发射的同时，进入具有变化的通道尺寸和形状的微流体芯片 背景减少和S/N增加。评估标准：（1）检测限（LOD）为1 CFU，（2）a 250μl的S/N，S/N，（3）填充时间小于30分钟。 一旦在第1阶段验证可行性后，这项工作就会通过实施来实现商业化的清晰途径 对Tracer-QC平台的测试。这将使宠物制造商在 符合所有合规标准的单个自动化系统。这项工作可以远远超出宠物 复合药房和其他小规模制造商，提供实验的简化 设置，生产的程序变化和调节性更改以提高患者安全性。