Droplet Digital PCR for absolute quantitation of viral-based biologics

点击翻译按钮获取中文摘要

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.