	US 12,234,277 B2
	Hydrophobic interaction chromatography for viral clearance
John Ruppino, Boonton, NJ (US); John Mattila, Nyack, NY (US); and Robert Stairs, Wellborn, FL (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Jan. 15, 2021, as Appl. No. 17/150,973.
Claims priority of provisional application 62/962,506, filed on Jan. 17, 2020.
Prior Publication US 2021/0221840 A1, Jul. 22, 2021
Int. Cl. C07K 16/06 (2006.01); A61K 39/395 (2006.01); C07K 1/20 (2006.01); C12Q 1/70 (2006.01); G01N 30/06 (2006.01); G01N 30/34 (2006.01)

CPC C07K 16/065 (2013.01) [C07K 1/20 (2013.01); C12Q 1/70 (2013.01); G01N 30/06 (2013.01); G01N 30/34 (2013.01)]

24 Claims

1. A method of purifying an antibody from a sample comprising one or more impurities including viral particles, the method comprising the steps of:

providing the sample comprising the antibody produced in a host-cell;

adding a sodium citrate buffer to the sample to produce a loading sample, wherein a concentration of the sodium citrate buffer is from about 10 mM to about 200 mM, and wherein adding the sodium citrate adjusts a hydrophobicity of the viral particles in the loading sample;

adjusting a pH of the loading sample to a range of from about 4.2 to about 8.0 by adjusting an amount of sodium citrate buffer;

loading the loading sample to a hydrophobic interaction chromatography (HIC) column to produce an HIC treated sample, wherein a concentration of the antibody in the loading sample is from about 40 g/L to about 200 g/L;

collecting the HIC treated sample; and

measuring a presence of viral genomic copies or viral particles in the collected HIC treated sample.