gRNA Analysis: Where's the Best Place to Start?

[kirstencraven]

There is a great deal of information in the public domain about the analysis of oligonucleotides by liquid chromatography-mass spectrometry (LC-MS). Its ability to measure multiple parameters, such as characterization and process control in a single analysis make it an appealing technique from early stage development to QC for biotherapeutics.

The complexity of the sample can make method development, system setup, and optimization tricky. Researching the analytical options and best practices can be frustrating – even when it’s a task you enjoy.

If you are analyzing gRNA or about to start on this journey, this paper published in the Journal of Chromatography A might be of interest, and save you some time. It talks in detail about the development of an ion pairing, reversed-phase liquid chromatography-mass spectroscopy (IR RPLC) method for the characterization of gRNA. This demonstrates its suitability for stability, characterization, and process control checks.

The authors found the Thermo Scientific™ DNAPac™ RP HPLC Column performed the best of the columns evaluated, delivering high resolution and separation of key compounds. They concluded these benefits were the result of the wide-pore particles used in the column — a proud and rewarding moment for our R&D department, who developed this highly robust column.

The LC-MS analysis of an oligonucleotide isn’t a task that is completed in isolation, and Thermo Fisher Scientific can help from the beginning to the end of your process. We also have preparative columns available with the same column chemistry, providing a simple and convenient end-to-end solution. Our preparative HPLC columns allow for easy, reliable and robust scale-up from analytical to preparative HPLC, even the most difficult analytes.

More information about all of the oligonucleotide HPLC columns available from Thermo Scientific is available in this brochure. Good luck with your method development and analysis!