US 12,351,612 B2
Method for preparing recombinant human nerve growth factor
Zhenping Zhu, Shanghai (CN); Haomin Huang, Shanghai (CN); and Jianguo Xiao, Shanghai (CN)
Assigned to SUNSHINE GUOJIAN PHARMACEUTICAL (SHANGHAI) CO., LTD., Shanghai (CN)
Appl. No. 17/787,558
Filed by SUNSHINE GUOJIAN PHARMACEUTICAL (SHANGHAI) CO., LTD., Shanghai (CN)
PCT Filed Nov. 3, 2020, PCT No. PCT/CN2020/126147
§ 371(c)(1), (2) Date Jun. 20, 2022,
PCT Pub. No. WO2021/120900, PCT Pub. Date Jun. 24, 2021.
Claims priority of application No. 201911325340.0 (CN), filed on Dec. 20, 2019.
Prior Publication US 2023/0037311 A1, Feb. 9, 2023
Int. Cl. C07K 14/48 (2006.01); C07K 1/14 (2006.01); C07K 1/20 (2006.01); C07K 1/30 (2006.01); C07K 1/34 (2006.01); C07K 1/36 (2006.01); C12N 15/70 (2006.01); C12P 21/02 (2006.01)
CPC C07K 14/48 (2013.01) [C07K 1/20 (2013.01); C07K 1/30 (2013.01); C07K 1/34 (2013.01); C07K 1/36 (2013.01); C12N 15/70 (2013.01); C12P 21/02 (2013.01)] 12 Claims
 
1. A method for preparing a purified preparation of recombinant human nerve growth factor comprising:
(a) expressing inclusion bodies of the recombinant human nerve growth factor in E. coli, and isolating the inclusion bodies from the E. coli;
(b) washing the inclusion bodies with an inclusion body washing solution;
(c) dissolving and denaturing the inclusion bodies by addition of a denaturing solution;
(d) renaturing the denatured inclusion bodies by addition of a renaturing solution and a buffer; and
(e) purifying the recombinant human nerve growth factor from the inclusion bodies;
wherein ammonium bicarbonate is used as the buffer in the step (d).