Amphastar Nanjing Pharmaceuticals Inc.

Huaiyan Cui

Song Chen

Haoning Zhang

Cheng Li

Chuangen Tang

Shangshu Pan

Xiaorui Liu

Jeffrey Ding

Provided are a novel pro-insulin aspart structure design and a method for preparing the pro-insulin aspart. The method comprises designing the pro-insulin aspart sequence, constructing a recombinant insulin aspart engineered bacteria, inducing expression of an insulin fusion protein as an inclusion body in the engineered bacteria, and extracting the insulin fusion protein from the engineered bacteria by means of denaturing, renaturing enzymatic digestion, and separation purification operations. By modifying the recombinant leading peptide and C-peptide sequences, the method avoids the need to include a cyanogen bromide cleavage step. In some embodiments, the C-peptide is replaced by a single amino acid which tends to reduce quality loss associated with enzymatic digestion.

Pro-insulin aspart structure and method for preparing insulin aspart

The invention belongs to the technical field of polypeptide preparation methods, and in particular relates to a preparation method of a liraglutide intermediate polypeptide GLP-1 (7-37). In the preparation method, main steps include constructing recombinant liraglutide engineered bacteria viato induce expression of a liraglutide intermediate fusion protein in the form of inclusion bodies, and performing denaturation, renaturation, enzyme digestion, separation and purification to obtain the liraglutide intermediate polypeptide GLP-1 (7-37). The invention alters expression pattern into the expression of the intracellular insoluble inclusion bodies by changing a signal peptide of the recombinant sequence to increase significantly expression level. The liraglutide intermediate polypeptide prepared by the invention has a purity up to 87% or more and a yield of more than 85%.

Method for preparing liraglutide intermediate polypeptide

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Jiangsu, China

Huaiyan Cui