Droplet Digital PCR for absolute quantitation of viral-based biologics

需要一种滴状数字聚合酶链式反应(DdPCR)系统来可靠和重复性地定量病毒疫苗和用于基因传递的病毒载体的细胞培养产品中的病毒产物的绝对浓度。DdPRC系统也将被用来描述干细胞培养中的稀有细胞群。在病毒载体和疫苗的生物处理实验室，麦吉尔大学生物工程系收购了Biorad QX200™和AutoDG™AutoDroplet Digital™聚合酶链式反应系统，大大节省了操作时间和成本。DdPCR可以在比目前可能的水平更低的水平上检测病毒，最大限度地减少珍贵样本的消耗，并更好地比较结果。DdPCR的大部分是自动化的，不需要参考标准，从而在更短的时间内提高了研究结论的统计可靠性，并降低了操作成本，因为不需要参考标准材料。这些设备对于申请者的研究项目更快、更有效地交付NSERC资助的CRC-1和Discovery项目以及NSERC资助的战略项目至关重要，这些项目以病毒载体和病毒疫苗的生物过程科学和技术为重点。化学工程系干细胞生物处理实验室的成员也将积极使用该设备，以量化稀有干细胞和干细胞衍生群体的基因表达，以实现更高质量的结果交付，并加速共同申请者基于干细胞的项目的发现。总体而言，ddPCR作为一种先进的过程分析技术的实施将加快疫苗和应用病毒学的研究和开发，并将产生新的发现，并加快与行业合作伙伴的研究成果的转化，并增加影响。申请人来自新成立的生物工程系的同事分享了对这种设备的需求，以可靠地量化他们的研究活动中涉及的核酸元素。随着目前和未来NSERC资助的项目和其他项目中研究的病毒、细胞培养和干细胞衍生的产品数量的增加，病毒基因组滴度测定设备的高频使用将扩大，机构合作伙伴将吸引更多的用户，重要的是培训更多的研究生了解这一核心分析技术。

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.