US 12,461,109 B2
Vitro prediction of in vivo half-life
Thomas Emrich, Iffeldorf (DE); Hubert Kettenberger, Munich (DE); Tilman Schlothauer, Penzberg (DE); and Angela Schoch, Munich (DE)
Assigned to Hoffmann-La Roche Inc.
Filed by Hoffmann-La Roche Inc., Little Falls, NJ (US)
Filed on Jan. 26, 2021, as Appl. No. 17/158,431.
Application 17/158,431 is a continuation of application No. 15/271,091, filed on Sep. 20, 2016, abandoned.
Application 15/271,091 is a continuation of application No. PCT/EP2015/055482, filed on Mar. 17, 2015.
Claims priority of application No. 14161103 (EP), filed on Mar. 21, 2014; and application No. 14165987 (EP), filed on Apr. 25, 2014.
Prior Publication US 2021/0190795 A1, Jun. 24, 2021
Int. Cl. G01N 33/68 (2006.01); B01D 15/16 (2006.01); B01D 15/38 (2006.01); C07K 16/00 (2006.01)
CPC G01N 33/6854 (2013.01) [B01D 15/168 (2013.01); B01D 15/3809 (2013.01); C07K 16/00 (2013.01); C07K 2317/52 (2013.01); C07K 2317/94 (2013.01)] 4 Claims
 
1. A method for selecting a variant antibody of a parent antibody, comprising the steps of:
determining a first retention time of the variant antibody and the parent antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a first salt concentration, and determining a second retention time of the variant antibody on an FcRn affinity chromatography column with the positive linear pH gradient elution in the presence of a second salt concentration, wherein:
the method is for selecting the variant antibody of the IgG1, IgG3 or IgG4 subclass that has a relative in vivo half-life that is increased compared to the parent antibody of the IgG1, IgG3 or IgG4 subclass, by selecting a variant antibody that has a first retention time that is longer than the first retention time of the parent antibody, and a first retention time that is substantially the same as the second retention time,
wherein the positive linear pH gradient is from about pH 5.5 to about pH 8.8; the salt is sodium chloride; and
wherein the first salt concentration is about 140 mM; and wherein the second salt concentration is about 400 mM.