High-Yield Biosynthesis Process for Producing Insulin Lispro.

Abstract

Insulin lispro, a fast-acting insulin analogue, is widely used for the treatment of diabetes due to its rapid onset and high flexibility. This study established a three-step chromatography purification process, L fermentation broth can obtain 475 mg/L insulin lispro. Through a multi-step process involving fermentation, recovery of inclusion bodies, renaturation, enzymatic digestion, and chromatographic purification, a high-purity insulin lispro product was obtained. The final product achieved a purity of 99.7%. The high molecular weight protein was ≤ 0.1%, zinc content was 0.41%, host cell protein contamination was 2.31 ng/mg, and residual DNA was 1.57 ng/mg. Analytical results, including retention time, molecular weight, and peptide mapping, were consistent with theoretical expectations and matched the control. This purification process provides a streamlined and cost-effective method for large-scale production of insulin lispro, supporting further development of recombinant insulin analogs and offering high-quality active pharmaceutical ingredients for pharmaceutical formulations.