Methods for purification of messenger RNA
First Claim
1. A method of purifying messenger RNA (mRNA), comprising(a) providing an impure preparation comprising in vitro synthesized mRNA and prematurely aborted RNA sequences,(b) treating the impure preparation with a denaturing agent under conditions that facilitate separation of the prematurely aborted RNA sequences from the mRNA;
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wherein purified mRNA from step (c) is substantially free of prematurely aborted RNA sequences and has an integrity greater than 95%.
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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 mRNA to a denaturing condition, and purifying the mRNA 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.
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Citations
26 Claims
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1. A method of purifying messenger RNA (mRNA), comprising
(a) providing an impure preparation comprising in vitro synthesized mRNA and prematurely aborted RNA sequences, (b) treating the impure preparation with a denaturing agent under conditions that facilitate separation of the prematurely aborted RNA sequences from the mRNA; - and
(c) subjecting the treated impure preparation from step (b) to tangential flow filtration, thereby purifying mRNA; wherein purified mRNA from step (c) is substantially free of prematurely aborted RNA sequences and has an integrity greater than 95%. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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Specification