Nucleotides are needed in many different areas, for example as building blocks for PCR and next generation sequencing, as building blocks for the selection of aptamers, or increasingly in therapy as prodrug agents (e.g. blockbuster agent sofosbuvir for the treatment of hepatitis C infections). The enzymatic synthesis of nucleotides, in contrast to chemical synthesis routes, does not require toxic reactants, expensive catalysts or toxic solvents and is therefore the method of the future.
However, bioengineered nucleotides are difficult and cost-inefficient to purify. High purity could now be achieved highly selectively and efficiently by ion exchange chromatography (IEX) followed by reverse phase chromatography (RPC). If these methods are coupled in an integrated counter current chromatography (iCCC), a highly productive and resource-saving technique is created.
In a ZIM-funded collaborative project between the Molnar Institute and Prof. Neubauer's group, the complex purification process consisting of IEX and RPC-iCCC as well as software for the iCCC will be developed. The process will be transferable to other small molecules.