	US 11,912,982 B2
	Methods for HPLC analysis
William Issa, Dedham, MA (US); and Meredith Packer, Waltham, MA (US)
Assigned to ModernaTX, Inc., Cambridge, MA (US)
Appl. No. 16/639,305
Filed by ModernaTX, Inc., Cambridge, MA (US)
PCT Filed Aug. 17, 2018, PCT No. PCT/US2018/046993 § 371(c)(1), (2) Date Feb. 14, 2020, PCT Pub. No. WO2019/036685, PCT Pub. Date Feb. 21, 2019.
Claims priority of provisional application 62/547,647, filed on Aug. 18, 2017.
Prior Publication US 2021/0163919 A1, Jun. 3, 2021
Int. Cl. C12N 15/10 (2006.01); G01N 30/02 (2006.01); G01N 30/88 (2006.01)

CPC C12N 15/101 (2013.01) [G01N 30/02 (2013.01); G01N 2030/8827 (2013.01); G01N 2496/00 (2013.01)]

20 Claims

1. A method for separating a nucleic acid having a hydrophobic portion from a mixture comprising one or more additional nucleic acids or impurities, the method comprising:

(i) contacting a hydrophobic stationary phase of a reverse phase chromatography column with the mixture; and

(ii) eluting the nucleic acid having a hydrophobic portion with a mobile phase, wherein the mobile phase: (a) comprises an ion pairing agent selected from (i) Tris, (ii) inorganic cations, and (iii) biological buffers, and (b) lacks triethylammonium salts, tetrabutylammonium salts, hexylammonium salts, and dibutylammonium salts; such that the nucleic acid having a hydrophobic portion traverses the column with a retention time that is different than the one or more additional nucleic acids or impurities of the mixture.