An eye on your nucleic acids with ÄKTA pure™ T system

While the world tried to understand and battle the COVID-19 pandemic. Scientists across the globe worked diligently on developing new vaccines and treatments. Ultimately mRNA vaccines showed the greatest efficacy in generating immunity against the virus and its variants. These mRNA vaccines were not only more efficient, they were faster and easier to manufacture than traditional vaccines. In less than a year after obtaining the genomic sequence of SARS CoV-2, the first COVID-19 vaccine was approved for emergency use by the FDA (Food and Drug Administration). Since then, mRNA vaccines for new strains have been developed and there are several ongoing clinical trials investigating their use for other types of infectious diseases, certain types of cancers, and a precursor for specific immunomodulatory proteins to treat chronic diseases (1, 2).

In vitro transcribed mRNA is not the only nucleic acid being investigated for its use as a therapeutic. Oligonucleotides (oligos), shorter strands of DNA and RNA, are also of interest for their therapeutic actions. Synthetic oligos have been used for decades in common molecular biology techniques like cloning and sequencing, and are now being investigated and used for more advanced applications like reagents in diagnostic kits and as gene silencing drugs for a range of different chronic diseases.

Purification and quality assessment of nucleic acids are indisputably important steps, as the quality of the sample will affect performance in later applications. Large amounts of pure sample is needed. Depending on the target, different nucleic acid characteristics can be obtained through several different methods like organic solvent extractions, precipitations, and magnetic particles.  Chromatography is gradually replacing these conventional methods because it can also remove templates, incomplete products, residual proteins, and impurities. In contrast to proteins, which absorb UV at 280 nm, nucleic acids absorb at 260 nm. Constant monitoring at this wavelength is preferred throughout a purification run with chromatography. The ratio of absorbance at 260 and 280 nm can also give you insights into the purity of a sample. A ratio of around 1.8 indicates pure DNA, a ratio of around 2.0 would indicate pure RNA, and a ratio of a pure protein typically has a value around 0.6.

We want to support our customers’ goals to generate data and insights that ultimately support the development of new types of therapies. Therefore, we developed the ÄKTA pure™ T system equipped with a fixed dual wavelength monitor, for real time and simultaneous measurement of 260 and 280 nm.  ÄKTA pure™ T system has all the features of our standard ÄKTA pure™ system, and also simplifies purification for a wider range of biomolecules like free DNA and RNA, or in complex with a protein. Together with UNICORN™ software, the ÄKTA pure™ T system generates a curve showing absorbance for 260 and 280 nm wavelengths for easy monitoring and calculation of the 260/280 ratio for your entire run as an initial assessment of sample purity.

So, what are you waiting for? Add your favorite resin to the ÄKTA pure™ T system and load your sample—to start keeping a close eye on your nucleic acids.