The modification of high-performance liquid chromatography (HPLC) parameters results in a better purification method for oligonucleotide-A. A method for optimizing the purification of an oligonucleotide-A on a reverse-phase C18 column using various experimental parameters such as buffer, concentration, and pH is developed. HPLC, UV/Vis, LC/MS, and lyophilization were used to purify oligonucleotide-A. Chromatographic data were collected using a semi-prep HPLC system, quantified using ultraviolet-visible spectrophotometry (UV-Vis), and confirmed using liquid-chromatography-mass spectrometry (LC-MS). The best possible optimized parameters found to achieve a 93.0% purity and an 84.0% yield are 40mM triethylammonium bicarbonate (TEAB) buffer with pH 7, which is about 6.0 and 5.0% higher than the reported method purity of 87.0% and yield of 80.0%, respectively. On the contrary, the worst possible optimized parameters that resulted in the lowest purity (84.0%) and yield (69.0%) are 10mM ammonium acetate with pH 7. A simple method for purifying oligonucleotide-A by HPLC has been optimized. When compared to previous methods, the purity increased by 6.0% and the yield increased by 5.0%.

Optimization of HPLC parameters for purification of oligonuceleotide-A

ACS Spring 2022

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