US 11,053,283 B2
Chromatography method
Jean-Luc Maloisel, Uppsala (SE); Ola Lind, Uppsala (SE); Bjorn Noren, Uppsala (SE); and Ronnie Palmgren, Uppsala (SE)
Assigned to Cytiva BioProcess R&D AB, Uppsala (SE)
Filed by Cytiva BioProcess R&D AB, Uppsala (SE)
Filed on Apr. 17, 2019, as Appl. No. 16/386,628.
Application 16/386,628 is a continuation of application No. 15/308,327, granted, now 10,266,563, issued on Apr. 23, 2019, previously published as PCT/EP2015/062500, filed on Jun. 4, 2015.
Claims priority of application No. 1450779-2 (SE), filed on Jun. 24, 2014.
Prior Publication US 2019/0241608 A1, Aug. 8, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 1/22 (2006.01); C07K 16/00 (2006.01); C07K 16/06 (2006.01); C07K 1/14 (2006.01)
CPC C07K 1/22 (2013.01) [C07K 16/00 (2013.01); C07K 16/065 (2013.01); C07K 1/145 (2013.01); C07K 2317/55 (2013.01)] 19 Claims
 
1. A method of making affinity chromatography beads suitable for purification of immunoglobulin containing proteins or parts thereof, from contaminants that are larger than immunoglobulin containing proteins, the method comprising:
providing allylated chromatography beads having an allylated outer layer and an allylated inner core;
inactivating the allylated outer layer of the allylated chromatography beads to form a lid consisting of agarose modified with hydrolyzed allyl groups and having a thickness of 1-8 μm;
immobilizing one or more affinity ligands to the allylated inner core of the allylated chromatography beads to form the affinity chromatography beads;
wherein the lid and the allylated inner core of the allylated chromatography beads have a defined pore size corresponding to KD of 0.1 to 1.