Plasmid DNA for vaccine and gene therapy research

The demand for high quality plasmid DNA has increased vastly in response to rapid advances in its use in gene therapy and vaccine production. New applications like mRNA based vaccines or gene therapy treatment all require plasmid DNA as an important starting material.

Plasmid DNA is, for example, an important template for mRNA vaccine production as it is the blueprint for the production of the mRNA.

Plasmids can also be used for direct injections into organisms, or as a tool to modify cells in vitro before they are injected into the organism.

NucleoBond PC EF - For purification of endotoxin-free plasmid DNA

A particularly challenging aspect of such procedures is the presence of endotoxins - Endotoxins are residual components of bacterial cell walls that can trigger an inflammatory response and dramatically decrease the efficiency of transfection in addition to distorting any results obtained from experiments.

Endotoxin-free plasmid is a crucial requirement for transfections and gene therapy applications. For this reason, MACHEREY-NAGEL created a proprietary technique for reducing endotoxins to extremely low levels. Learn more about our NucleoBond PC 10000 kits in this workflow video:

How do endotoxins affect downstream applications?

Endotoxins have cytotoxic effects and negatively influence cell viability and transfection efficiency. Additionally, endotoxins are known to influence gene expression in cell cultures, leading to false results in gene expression analyses. Endotoxin levels are measured by a standardized test, and the measurement unit is EU (endotoxin unit). 

How to pick the right product? 

The plasmid purification product that best fits your needs will depend upon your downstream application. Routine molecular biology applications are not influenced by the presence of endotoxins. However, manipulation of standard eukaryotic cell lines such as HeLa or HEK cells require a higher purity. For these applications, we recommend our transfection-grade plasmid DNA isolation kits with endotoxin levels between 1–50 EU/μg DNA. For highly sensitive applications, we recommend our endotoxin-free kits. These applications include the transfection of precious cell lines, such as primary cells, stem cells or cells growing in low numbers. Our endotoxin-free products enable the isolation of plasmid DNA with endotoxin levels below 0.1 EU/μg DNA.