Abstract
<title>Abstract</title> <p>RNaseR, an exoribonuclease, selectively targets linear RNA from the 3’-5’ direction in the presence of other RNA species, and process intermediates. To date, no comparative studies have evaluated the digestion efficiencies of RNaseRs procured from different vendors. In this study, we assessed the performance of RNaseRs from four vendors on RNA vaccine drug substance (DS). The results revealed significant variations in digestion efficiencies among the enzymes evaluated. Furthermore, we investigated the effects of digestion incubation time and substrate-to-enzyme ratios, as well as comparing various buffer compositions (including vendor-provided buffers and our in-house LiCl-based buffer formulation) on RNaseR digestion activity. Our findings indicate that specific buffer matrices can significantly enhance RNaseR digestion, highlighting the importance of buffer optimization. In conclusion, this study offers valuable insights into the performance of RNaseRs from different vendors and underscores the need for optimized buffer conditions to improve the degradation of linear RNA species.</p>