By Nina Mencin, Urh Černigoj, Sebastijan Peljhan, Špela Peršič, Pete Gagnon, Aleš Štrancar
The increasing demand for messenger RNA (mRNA) as a therapeutic product requires larger production scales and more efficient extraction techniques. Here the fast and efficient way to purify polyadenylated mRNA using affinity chromatography on the CIMmultus® Oligo dT column is presented.
The poly-adenylated tail of mRNA interacts with covalently bound oligo dT ligands in high-salt loading conditions, where electrostatic repulsion between negatively charged backbones of both mRNA and oligo dT are reduced, and H-bonding in the T-A base pair is emphasized. High salt concentration also screens out attractive electrostatic interactions between mRNA and other components in the process sample, thus facilitating an aggregate reduction in purified products (Figure 1).