HPLC Purification of IVT mRNA
To date, bacteriophage-derived RNA polymerases have served as key enzymes for the efficient production of in vitro-transcribed (IVT) mRNA. Large quantities of RNA can be easily yielded by in vitro transcription from DNA templates using phage RNA polymerase. However, impurities are generated during the reaction due to various unwanted activities of the polymerases. Besides, remaining reagents, such as DNA templates, unreacted nucleotides, and RNA polymerase, all need to be eliminated. Here, we provide an efficient and scalable HPLC purification service to remove multiple contaminants from IVT mRNA. Using our platform, we can translate HPLC-purified mRNA at much greater levels to be used widely in primary cells, in vivo and among others.
Advantage of HPLC purification
The HPLC method can be utilized to isolate synthesized mRNA from the remaining reagents via size-exclusion or ion-exchange columns, thereby removing smaller or larger by-products, such as double-stranded RNA, abortive transcripts and so on. In addition, chromatography offers a closed system to minimize the risk of cross-contamination or exposure to RNase degradation. As an easily scalable platform, the HPLC method can produce large amounts of RNA necessary for therapeutic applications and can be completed quickly and efficiently.
For large-scale purification and isolation, high performance liquid chromatography (HPLC) is most often used. Besides the HPLC purification service, we also offer multiple complementary services to ensure that high-quality IVT mRNA will be delivered to our customers.
HPLC purification of IVT mRNA
Our HPLC purification service is based on a robust and optimal workflow, including determining the separation goals, choosing detector and detector settings, optimizing separation conditions and checking for problems. To acquire better HPLC purification experience, before applying the HPLC column, we adopt LiCl sedimentation, a standard method of initial purification to treat the in vitro transcription reaction. It is necessary to use a standard purification before HPLC purification to identify the overloading of the HPLC column. In addition, to achieve the best purification results, we strictly abide by standard HPLC protocols for storage and maintenance of the HPLC columns and system.
Our RNA analysis services contain RNA translation, RNA immunogenicity analyses, and dsRNA dot blot. (dsRNA is a major contaminant in IVT mRNA synthetic reaction system that can activate RNA sensors to inhibit protein translation directly or indirectly.) It is necessary to determine the contaminant after HPLC purification.
To meet the particular needs of global customers, Creative Biogene provides the high-purity and high-yield IVT mRNA based on advanced and robust technology platforms. We believe that our service will accelerate the progress of your project. Please don't hesitate to contact us to request an IVT mRNA purification service.