Purification and Evaluation of the Biological Activity of Recombinant Mouse Prolactin.

Abstract

Prolactin, a hormone secreted by lactotroph cells in the anterior pituitary lobe, plays a crucial role in vascular growth and immune homeostasis. In this study, we evaluated the functional activity of recombinant mouse prolactin produced using Escherichia coli. By employing a maltose-binding protein tag and TEV protease cleavage, we enhanced protein solubility and stability while ensuring high bioactivity. The recombinant prolactin was purified to over 99% purity through a streamlined two-step chromatography process, as confirmed by SDS-PAGE and Western blot analyses. Cell-based assays verified that the purified prolactin retained its biological activity, indirectly demonstrating its structural and functional stability. Additionally, the cost-effectiveness and simplicity of this method were highlighted by the reusability of the affinity column, reducing purification costs compared to conventional methods. This study provides a simple purification method for obtaining activated recombinant mouse prolactin, suggesting its potential use in biological and biochemical research.