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US 11,976,272 B2Granted
Methods for purification of messenger RNA
AssigneeTranslate Bio, Inc.
InventorsJonathan Abysalh, Daniel Crawford, Frank DeRosa, Shrirang Karve, Anusha Dias, Michael Heartlein
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Abstract
The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, steps of forming an mRNA slurry, stirring the slurry, and vacuum or pressure filtering the slurry.
Claims21
A method of purifying at least about 1 gram of in vitro transcribed mRNA, comprising steps of:
- providing a solution comprising the in vitro transcribed mRNA;
- adding one or more agents that promote precipitation of the in vitro transcribed mRNA, thereby obtaining a slurry;
- stirring the slurry prior to and/or while providing pressure to the slurry and/or a vacuum to the slurry sufficient to direct the slurry's mother liquor through a filter, thereby obtaining a precipitate-containing composition;
- and
- washing the precipitate-containing composition, thereby yielding a purified mRNA precipitate;
- wherein the slurry comprises at least one dispersant;
- wherein at least 1 gram of purified mRNA is obtained, and wherein the purified mRNA has reduced impurity of enzyme reagents used in in vitro transcription as compared to the otherwise same mRNA purified by tangential flow filtration.
4 more elements
A method for purifying at least about 1 gram of in vitro transcribed mRNA, comprising steps of:
- providing a solution comprising the in vitro transcribed mRNA;
- adding one or more agents that promote precipitation of the in vitro transcribed mRNA, thereby obtaining a slurry;
- stirring the slurry prior to and/or while providing pressure to the slurry and/or a vacuum to the slurry sufficient to direct the slurry's mother liquor through a filter, thereby obtaining a precipitate-containing composition;
- and
- washing the precipitate-containing composition, thereby yielding a purified mRNA precipitate;
- wherein the purified mRNA has reduced impurity of enzyme reagents used in in vitro transcription as compared to the otherwise same mRNA purified by tangential flow filtration;
- wherein the total purified mRNA is recovered in an amount that results in a yield of at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95% or the total purified mRNA is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis;
- wherein the slurry comprises at least one dispersant;
- and wherein at least 1 gram of purified mRNA is obtained.
6 more elements
Dependencies are inferred from the claim text.