Effect of Mg 2+ Ion Concentration on IVT Reaction Kinetics Determined by Novel Rapid Analytical HPLC Assay

The IVT reaction is one of the most expensive steps in mRNA production process and its optimization to reach high mRNA yield is of key importance. Standard mRNA quantification techniques like absorbance and fluorescence based assays are time consuming and cannot be performed at line as the IVT reaction progresses. In addition, other reaction components like nucleotides and pDNA interfere in the analytical results and reduce the method’s accuracy.

A new approach shown here uses CIMac PrimaS analytical HPLC column to separate and quantify several key IVT components with a very short run time, enabling fast “at line” tracking.

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