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Single-Use Based BioProduction Process for Manufacturing of High Quality Recombinant Enzymes for mRNA and DNA Workflows
EP31092
Poster Title: Single-Use Based BioProduction Process for Manufacturing of High Quality Recombinant Enzymes for mRNA and DNA Workflows
Submitted on 16 Dec 2019
Author(s): Darius Kavaliauskas, Viktorija Vitkovskė, Ramunė Leipuvienė, Juozas Šiurkus
Affiliations: Thermo Fisher Scientific Baltics
Poster Views: 196
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Poster Information
Abstract: Molecular research and therapy relies heavily on tests utilizing
polymerase chain reaction (PCR) and in vitro transcription
(IVT) technologies. PCR enables sensitive detection, specific
identification, and accurate quantification of nucleic acid
sequences and IVT is used to manufacture mRNA. To avoid
false positives and uncertainty in the assay results, it is crucial
that the reagents used in PCR and IVT applications be free of
foreign contaminants, such as DNA, RNA, HCP, endotoxins
originating from the manufacturing environment, human
operator, cross-contamination from shared equipment, or host
cells expressing recombinant proteins. To remove the risk of
these types of contamination, Thermo Fisher Scientific has
developed and implemented a new process for manufacturing
such enzymes. We use a closed system based on single-use
technology to drastically minimize the risk of contamination
inherent to the conventional manufacturing process. To help
ensure conformance to strict purity requirements, we subject
our DNA-free PCR reagents and IVT enzymes to stringent
quality tests. This combination of manufacturing systems and
quality testing delivers PCR reagents that are orders of
magnitude cleaner than other “DNA-free” reagents on the
market and IVT reagents that meet all quality and regulatory
requirements for therapeutic use.
Summary: Conventional PCR and IVT reagents, which have been shown
to contain contaminating DNA/RNA, fall short of being able to
provide the reliability required to detect low abundance
DNA/RNA targets. To support developers of DNA based assay
and IVT kits, Thermo Fisher Scientific is the first to offer
DNA/RNA-free PCR and IVT reagents manufactured using a
closed single-use system technology.
References: 1. Salter SJ, Cox MJ, Turek EM et al. (2014) Reagent and
laboratory contamination can critically impact sequence-based
microbiome analyses. BMC Biology 12:87.
2. Konstantinov KB, Cooney CL (2015) White paper on
continuous bioprocessing. J Pharm Sci 104:813–20.
3. Czurda S, Smelik S, Preuner-Stix S et al. (2016)
Occurrence of fungal DNA contamination in PCR reagents:
approaches to control and decontamination. J Clin Microbiol
54:148–152
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