US 12,247,082 B2
Highly efficient anti-TFPI antibody composition
Min Jung Kim, Yongin-si (KR); Geun Hye Yeo, Yongin-si (KR); Haeng Eun Song, Yongin-si (KR); Ji Yoon Park, Yongin-si (KR); and Yuna Kim, Yongin-si (KR)
Assigned to GREEN CROSS CORPORATION, Yongin (KR)
Appl. No. 17/276,276
Filed by GREEN CROSS CORPORATION, Yongin-si (KR)
PCT Filed Sep. 6, 2019, PCT No. PCT/KR2019/011582
§ 371(c)(1), (2) Date Mar. 15, 2021,
PCT Pub. No. WO2020/060087, PCT Pub. Date Mar. 26, 2020.
Claims priority of application No. 10-2018-0113974 (KR), filed on Sep. 21, 2018.
Prior Publication US 2022/0025067 A1, Jan. 27, 2022
Int. Cl. C07K 16/28 (2006.01); C07K 1/16 (2006.01); C07K 1/22 (2006.01); C07K 1/34 (2006.01); C07K 1/36 (2006.01); C07K 16/38 (2006.01)
CPC C07K 16/38 (2013.01) [C07K 1/165 (2013.01); C07K 1/22 (2013.01); C07K 1/34 (2013.01); C07K 1/36 (2013.01); C07K 2317/10 (2013.01); C07K 2317/92 (2013.01)] 8 Claims
 
1. A method of preparing a pharmaceutical composition comprising a purified anti-tissue factor pathway inhibitor (anti-TFPI) antibody, wherein a content of host cell protein (HCP) in the composition is less than 10.0 ng/mg, and a content of leachable protein A (LPA) in the composition is less than 1.0 ng/mg,
wherein the anti-TFPI antibody is an IgG4 antibody,
the method comprising
(a) a step of clarifying a culture containing the anti-TFPI antibody to obtain a clarified culture, wherein the clarified culture is subjected to following (b)-(g), in this order;
(b) an affinity chromatography process at pH of 3.0 to 3.8, which further comprises the steps of:
(b-1) neutralizing an eluate, which passed through an affinity chromatography column, in the range of pH 4.0 to 6.0; and
(b-2) leaving the neutralized eluate obtained in (b-1) to stand at a temperature of 15 to 25° C. for 8 to 16 hours;
(c) a viral inactivation process;
(d) an ultrafiltration and diafiltration process;
(e) a mixed-mode chromatography process;
(f) a nano-filtration process; and
(g) a concentration process,
wherein the mixed-mode chromatography process (e) comprises performing two or more plural mixed-mode chromatography processes using different types of column resin to remove the anti-TFPI antibody in a form of polymer.