Poster Presentation at the 11th Annual Meeting of the Nucleic Acids Therapeutics Society of Japan: “Development of Chromatographic Separation Methods for the Purification of Single-Guide RNA Therapeutics” (Presented in Collaboration with YMC CO., LTD.)
At the 11th Annual Meeting of the Nucleic Acids Therapeutics Society of Japan, held at Dejima Messe Nagasaki from July 7th to 9th, YMC Co., Ltd. delivered a poster presentation on our joint research. NIPPON SHOKUBAI CO., LTD. participated as a co-presenter, collaborating on the poster preparation and joint Q&A sessions. We are pleased to introduce the content published in the meeting’s abstract collection.
Presentation
| Title | Development of Chromatographic Separation Methods for the Purification of Single-Guide RNA Therapeutics |
| Affiliations | R&D DEPT., YMC CO., LTD., Health & Medical Business Division, NIPPON SHOKUBAI CO., LTD. |
Background
Demand for highly pure single-guide RNA (sgRNA) is increasing as its use in clinical applications grows. Ion-pair reversed-phase liquid chromatography (IP-RP-LC) using alkylamine ion-pair reagents and hexafluoroisopropanol (HFIP) has been widely used for high-resolution separation of long (approximately 100-mer) oligonucleotides, enabling the removal of closely related impurities from synthetic sgRNA. This method is, however, hardly applicable to industry-scale purification of long oligonucleotides due to the high cost and environmental burden of HFIP; thus, an alternative method is required.
Research Contents
This study aimed to develop an IP-RP-LC method for purifying phosphorothioate (PS)-modified sgRNA without using HFIP or other expensive additives. Previous research indicates that the addition of sodium phosphate buffer to alkylamine mobile phase solutions enhances the separation of long oligonucleotides. Here, we hypothesized that the following two factors are involved in this enhancement: 1) phosphate ions, which may prevent the adsorption of oligonucleotides in the LC system, including the column; 2) sodium ions in the mobile phase, which may stabilize the secondary structure of oligonucleotides. Our data show that the enhanced separation of oligonucleotides by the addition of phosphate buffer is due to sodium ions, rather than to phosphate ions. We observed that a similar enhancement was also caused by another potassium ion. By applying these findings, we established a novel method for separating 100-mer sgRNA from 5’ truncated products and phosphodiester (PO) impurities without using HFIP. This method offers a cost-effective and environmentally favorable solution for the industrial manufacturing of sgRNA drug substances.
Meeting Information
| Name | The 11th Annual Meeting of the Nucleic Acids Therapeutics Society of Japan |
| Meeting | July 7th (Tue) – July 9th (Thu), 2026 |
| Venue | Dejima Messe Nagasaki |
| Official website | https://www.natsj.jp/2026/natsj11 |