US 12,129,460 B2
Methods for ethanol-free mRNA purification
Eric Calvosa, Paris (FR); Nicolas Chaudet, Paris (FR); Arthur Leclercq, Paris (FR); and Alban Lepetitcolin, Paris (FR)
Assigned to SANOFI, Paris (FR)
Filed by SANOFI, Paris (FR)
Filed on Jul. 28, 2023, as Appl. No. 18/360,978.
Claims priority of application No. 22315175 (EP), filed on Jul. 29, 2022; and application No. 23173491 (EP), filed on May 15, 2023.
Prior Publication US 2024/0043826 A1, Feb. 8, 2024
Int. Cl. C12N 15/10 (2006.01)
CPC C12N 15/101 (2013.01) [C12N 15/1017 (2013.01)] 21 Claims
 
1. A method of purifying messenger RNA (mRNA), comprising:
(a) subjecting a preparation comprising in vitro synthesized mRNA to an enzymatic digestion with a proteinase;
(b) subjecting the preparation obtained from step (a) to a step of tangential flow filtration (TFF);
(c) subjecting the preparation obtained from step (b) to a step of capping;
(d) subjecting the preparation obtained from step (c) to a step of frontal filtration;
(e) subjecting the preparation obtained from step (d) to an oligo dT affinity chromatography; and
(f) subjecting the preparation obtained from step (e) to a step consisting of TFF, thereby purifying the mRNA.