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TFF in RNA workflow: Process optimization for RNA and LNP manufacturing | Poster Board #310
Date
March 25, 2025
The advent of messenger RNA (mRNA) vaccines for COVID-19 has revolutionized RNA-based genomic medicine and opened avenues for prophylactic as well as therapeutic applications in multiple disease areas. Apart from mRNA, self-amplifying/-replicating (saRNA) and circular RNA (cRNA) have emerged as new classes of RNA drug molecules.
While RNA and lipid nanoparticle (LNP) processing is still evolving, the industry seems to be converging on TFF technologies for concentration and buffer exchange given their cost-effectiveness, scalability, and proven track record in the biotech industry. Ultrafiltration and diafiltration (UF/DF) via tangential flow filtration (TFF) is an important unit operation in RNA manufacturing that is typically employed in three to four steps in RNA and LNP downstream processing. A longstanding question for TFF optimization of RNA and LNPs is filter format- whether to use a hollow fiber or flat sheet cassette. There is also a limited understanding of the impact of membrane chemistry for downstream processing of RNA and LNPs.
RNA and LNP UF/DF processes are typically run at ambient temperature wherein maintaining product quality and minimizing processing time are paramount. Understanding the importance of membrane chemistry and format (hollow fibers and flat sheet cassettes) in process optimization of TFF for mRNA and LNPs is imperative. Here we take a deep dive into TFF optimization for RNA and LNPs using 100 kDa molecular weight cut off membrane devices. Our data reveals how process conditions such as membrane chemistry and filter format can significantly impact process optimization for mRNA and LNPs. We show that a single filter can work across modalities (1 kilobase mRNA, 8.5 kilobase saRNA, LNP) and discuss the value in streamlining manufacturing stocking and operations. Finally, we share scalability data from bench- (93 cm2) to manufacturing-scale (0.5-1 m2) using mRNA and LNPs, discussing proper scaling factors and any offsets observed in the data.
While RNA and lipid nanoparticle (LNP) processing is still evolving, the industry seems to be converging on TFF technologies for concentration and buffer exchange given their cost-effectiveness, scalability, and proven track record in the biotech industry. Ultrafiltration and diafiltration (UF/DF) via tangential flow filtration (TFF) is an important unit operation in RNA manufacturing that is typically employed in three to four steps in RNA and LNP downstream processing. A longstanding question for TFF optimization of RNA and LNPs is filter format- whether to use a hollow fiber or flat sheet cassette. There is also a limited understanding of the impact of membrane chemistry for downstream processing of RNA and LNPs.
RNA and LNP UF/DF processes are typically run at ambient temperature wherein maintaining product quality and minimizing processing time are paramount. Understanding the importance of membrane chemistry and format (hollow fibers and flat sheet cassettes) in process optimization of TFF for mRNA and LNPs is imperative. Here we take a deep dive into TFF optimization for RNA and LNPs using 100 kDa molecular weight cut off membrane devices. Our data reveals how process conditions such as membrane chemistry and filter format can significantly impact process optimization for mRNA and LNPs. We show that a single filter can work across modalities (1 kilobase mRNA, 8.5 kilobase saRNA, LNP) and discuss the value in streamlining manufacturing stocking and operations. Finally, we share scalability data from bench- (93 cm2) to manufacturing-scale (0.5-1 m2) using mRNA and LNPs, discussing proper scaling factors and any offsets observed in the data.
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