公开(公告)号：EP4328316A1

公开(公告)日：2024-02-28

申请号：EP21940283.1

申请日：2021-05-31

申请人： Asymchem Laboratories (Tianjin) Co., Ltd

发明人： HONG, Hao ,  JAMES, Gage ,  XIAO, Yi ,  ZHANG, Na ,  JIAO, Xuecheng ,  ZHAO, Junqi ,  WANG, Lei ,  MENG, Xiangyu ,  ZHANG, Mujiao

IPC分类号： C12P21/02 ,  C12P21/06

摘要： Provided is a preparation method for a polypeptide. The preparation method includes the following steps: constructing an engineering strain for fusion-expressing a polypeptide gene with a Sumo tag, and inducing the engineering strain for the soluble expression of a polypeptide; obtaining a fusion protein containing a polypeptide precursor from the engineering strain by purification; cleaving the fusion protein containing the polypeptide precursor by a Ulp1 protease to remove the Sumo tag; and purifying a cleavage product of the Ulp1 protease by a method of acetonitrile combined with heating precipitation or a method of precipitation with hexafluoroisopropanol to obtain the polypeptide. By establishing an efficient technology based on soluble recombinant expression of a medicinal polypeptide or a precursor thereof and establishing a simple purification process based on one-step precipitation purification, further combined with HPLC refinement, a polypeptide purity of over 97% can be achieved.