Amphastar Nanjing Pharmaceuticals Inc.

Jeffrey Ding

Song Chen

Haoning Zhang

Cheng Li

Chuangen Tang

Xiaorui Liu

Shangshu Pan

Huaiyan Cui

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

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Jiangsu, China

Jeffrey Ding