Droplet Digital PCR for absolute quantitation of viral-based biologics

需要一种液滴数字PCR （ddPCR）系统来可靠地、可重复地定量病毒疫苗和基因传递病毒载体细胞培养产品中的病毒产物的绝对浓度。ddPRC系统也将用于表征干细胞培养中的稀有细胞群。麦吉尔大学生物工程系在病毒载体和疫苗的生物处理实验室中获得了ddPCR，如Biorad QX200™与AutoDG™AutoDroplet Digital™PCR系统，这大大节省了操作时间和成本。ddPCR可以在比目前可能的更低水平上检测病毒，最大限度地减少宝贵样品的消耗，并更好地比较结果。大部分ddPCR是自动化的，不需要参考标准，在更短的时间内提高了研究结论的统计可靠性，并且由于不需要参考标准材料，操作成本更低。该设备对于申请人的研究项目至关重要，可以更快、更有效地在nserc资助的CRC-1和发现项目以及nserc资助的战略项目中提供，以病毒载体和病毒疫苗的生物过程科学和技术为中心。该设备还将被化学工程系干细胞生物处理实验室的成员积极使用，以量化稀有干细胞和干细胞衍生群体的基因表达，以获得更高质量的结果，并加速共同申请人基于干细胞的项目的发现。总体而言，将ddPCR作为一种先进的过程分析技术加以实施，将加速疫苗和应用病毒学的研究与开发，并将产生新的发现，加速与工业伙伴将研究成果转化，并增加影响。申请人在新成立的生物工程系的同事也需要这种设备，以可靠地量化他们研究活动中涉及的核酸元素。随着目前和未来nserc资助的项目以及其他与机构合作伙伴合作的项目中研究的病毒、细胞培养和干细胞衍生产品的数量的增加，病毒基因组滴度测定设备的高频使用将会扩大，这些项目将吸引更多的用户，重要的是培训更多的研究生学习这一核心分析技术。

A droplet digital PCR (ddPCR) system is requested to reliably and reproducibly quantify the absolute concentrations of viral products in cell culture productions of viral vaccines and viral vectors for gene delivery. The ddPRC system will also be used to characterize rare cell population in stem cell cultures. The acquisition of ddPCR, such as the Biorad QX200™ with AutoDG™ AutoDroplet Digital™ PCR System in the bioprocessing of viral vectors and vaccines laboratory, Bioengineering Department at McGill University represents a significant time and cost saving on operations. ddPCR allows detection of virus at lower levels than is presently possible, minimal consumption of precious samples and better comparison of results. Much of the ddPCR is automated and reference standards are not required, improving the statistical reliability of research conclusions within shorter timelines and at lower costs of operation since no reference standard material is needed. The equipment is critical for the research programs of the applicant to deliver more rapidly and more effectively on NSERC-funded CRC-1 and Discovery programs as well as NSERC-funded strategic projects, pivoting on the bioprocess science and technology of viral vectors and viral vaccines. The equipment will also be actively used by members of the Stem Cell Bioprocessing Laboratory in the Department of Chemical Engineering in order to quantify gene expression by rare stem cell and stem cell-derived populations for higher quality results delivery and acceleration of discovery on stem cell-based projects of the co-applicant. Overall, the implementation of ddPCR as an advanced process analytical technology will accelerate the vaccine and applied virology research and development and will result in new findings and accelerated translation of research results with industrial partners and increased impact. The applicant’s colleagues from the newly created bioengineering department share the needs for such equipment to reliably quantify nucleic acid elements involved in their research activities. The high frequency use of the equipment for determination of viral genome titers will expand with the number of products derived from virus, cell culture and stem cells that are studied in the current and future NSERC-funded projects and other projects with institutional partners engaging more users and importantly training more graduate students on this core analytical technology.