Methods for purification of messenger RNA
AssigneeTranslate Bio, Inc.
InventorsJonathan Abysalh, Frank DeRosa, Jorel E. Vargas, Cameron M. Smith
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Abstract
The present invention provides, among other things, methods for purifying high quality messenger RNA (mRNA) suitable for clinical use, without using any caustic or flammable solvents. The present invention is, in part, based on surprising discovery that mRNA can be successfully purified by selective precipitation and washing without using ethanol while maintaining integrity and high purity of mRNA. Thus, the present invention provides an effective, reliable, and safer method of purifying RNA from large scale manufacturing process therapeutic applications without using any caustic or inflammable solvents.
Claims8
A method of purifying messenger RNA (mRNA) comprising:
- a) precipitating the mRNA in a suspension comprising at least a 1M salt solution and an amphiphilic polymer;
- b) capturing the mRNA on a filter;
- and
- c) washing the mRNA of step b) with a triethylene glycol monomethyl ether (MTEG) solution to obtain a purified mRNA composition substantially free of contaminants.
A method of purifying messenger RNA (mRNA) comprising:
- a) precipitating the mRNA in a guanidinium thiocyanate (GSCN) solution comprising MTEG;
- b) centrifuging the solution of step a) to create an mRNA pellet;
- c) resuspending the mRNA pellet in a buffer;
- d) capturing the mRNA on a filter;
- e) washing the mRNA of step d) with a MTEG solution;
- and
- f) solubilizing the washed mRNA of step e) to obtain an mRNA composition substantially free of contaminants.
Dependencies are inferred from the claim text.