US 12,084,702 B2
Methods for purification of messenger RNA
Joseph Parella, Lexington, MA (US); Kimberly Gillis, Lexington, MA (US); Jonathan Abysalh, Lexington, MA (US); Travis Jeannotte, Lexington, MA (US); Erik Held, Lexington, MA (US); Frank DeRosa, Lexington, MA (US); Michael Heartlein, Lexington, MA (US); and Rebecca Powell, Lexington, MA (US)
Assigned to TRANSLATE BIO, INC., Waltham, MA (US)
Filed by Translate Bio, Inc., Lexington, MA (US)
Filed on Oct. 11, 2021, as Appl. No. 17/450,506.
Application 17/450,506 is a continuation of application No. 16/551,340, filed on Aug. 26, 2019, granted, now 11,174,500.
Claims priority of provisional application 62/722,674, filed on Aug. 24, 2018.
Prior Publication US 2022/0170061 A1, Jun. 2, 2022
Int. Cl. C12P 19/34 (2006.01); B01D 25/00 (2006.01); B01D 37/00 (2006.01); C07H 1/06 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01); C12N 15/10 (2006.01); C12N 15/52 (2006.01)
CPC C12P 19/34 (2013.01) [B01D 25/00 (2013.01); B01D 37/00 (2013.01); C07H 1/06 (2013.01); C07H 21/02 (2013.01); C07H 21/04 (2013.01); C12N 15/1017 (2013.01); C12N 15/52 (2013.01); C12N 2330/30 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method for purifying messenger RNA (mRNA) manufactured by in vitro transcription (IVT) synthesis, the method comprising the steps of:
(a) precipitating mRNA from a solution comprising one or more contaminants from manufacturing the mRNA to provide a suspension comprising precipitated mRNA;
(b) subjecting the suspension comprising the precipitated mRNA to a depth filter comprising three or more filter layers of a single three-dimensional matrix, or three or more filter layers of different three-dimensional matrices, wherein each of the three or more layers has a pore size that is smaller than the pore size of the preceding layer in the direction of flow and wherein the precipitated mRNA is retained by the depth filter;
(c) dissolving the precipitated mRNA retained by the depth filter in step (b) thereby allowing purified mRNA to pass through the depth filter; and
(d) recovering the purified mRNA from step (c);
wherein at least 1 gram of mRNA is purified per batch and the depth filter is made of an inert material that allows capture of the precipitated mRNA without clogging or forming a gel layer.