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US 12,760,296 B2Granted
Methods for purification of messenger RNA
AssigneeTranslate Bio, Inc.
InventorsFrank DeRosa, Anusha Dias, Michael Heartlein, Shrirang Karve
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Abstract
The present invention provides, among other things, methods of purifying messenger RNA (mRNA) including the steps of subjecting an impure preparation comprising in vitro synthesized rnRNA to a denaturing condition, and purifying the rnRNA from the impure preparation from step (a) by tangential flow filtration, wherein the mRNA purified from step (b) is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis.
Claims23
A method of purifying messenger RNA (mRNA), comprising
- (a) providing an impure preparation comprising in vitro synthesized mRNA and enzyme reagents;
- (b) treating the impure preparation comprising in vitro synthesized mRNA with a denaturing agent under conditions that facilitate separation of the enzyme reagents from the mRNA;
- and
- (c) subjecting the treated impure preparation from step (b) to tangential flow filtration (TFF), thereby purifying the mRNA;
- wherein the mRNA purified from step (c) is substantially free of the enzyme reagents;
- wherein the mRNA is purified at a scale of 5 milligrams or greater per batch;
- and
- wherein the purified mRNA is suitable for administration as a pharmaceutical product.
5 more elements
Dependencies are inferred from the claim text.