Scalable multimodal weak anion exchange chromatographic purification for stable mRNA drug substance

Messenger RNA (mRNA) has emerged as a modality with immense therapeutic potential. Recent innovations in production process of mRNA call for procedures to isolate pure mRNA drug substance (DS) with high yield, high capacity, scalability, and compatibility with GMP production systems. Novel RNA modal...

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Bibliographic Details
Published in:Electrophoresis 2023-12, Vol.44 (24), p.1978-1988
Main Authors: Megušar, Polona, Miklavčič, Rok, Korenč, Matevž, Ličen, Jure, Vodopivec, Tina, Černigoj, Urh, Štrancar, Aleš, Sekirnik, Rok
Format: Article
Language:English
Subjects:
Anion exchanging
Chemical bonds
Chromatography
high‐performance liquid chromatography
Hydrogen bonding
in vitro transcription
Ion exchange
Monoclonal antibodies
mRNA
phosphate
Purification
Room temperature
Stability
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Summary:Messenger RNA (mRNA) has emerged as a modality with immense therapeutic potential. Recent innovations in production process of mRNA call for procedures to isolate pure mRNA drug substance (DS) with high yield, high capacity, scalability, and compatibility with GMP production systems. Novel RNA modalities, such as circular RNA (circRNA), have further driven the need for non‐affinity capture possibilities which are already widely used in the biopharmaceutical industry, for example, in monoclonal antibody processing. The principle that multimodal ion exchange/hydrogen bonding chromatography can be used to separate mRNA from in vitro transcription components has recently been demonstrated. Here, we apply and refine this approach to be suitable for scalable purification of multiple mRNA constructs with sufficient yields, purity, and stability, for use in mRNA production process. Binding capacity of the PrimaS‐modified monolithic chromatographic column for mRNA enabled up to 7 mg/mL product isolation in a single chromatographic run, with 98% recovery and room temperature stability of the eGFP mRNA demonstrated for up to 28 days. This approach is independent of construct size or the presence of polyadenylic acid tail and is applicable for capture of a wide variety of RNAs, including mRNA, self‐amplifying RNA, circRNA, and with optimization also smaller RNAs such as transfer RNA and others.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.202300106