Soluble Expression of Human Leukemia Inhibitory Factor with Protein Disulfide Isomerase in Escherichia coli and Its Simple Purification

摘要: Human leukemia inhibitory factor (hLIF) is a multifunctional cytokine that essential for maintaining the pluripotency of embryonic stem cells. hLIF may be also useful in aiding fertility through its effects on increasing implantation rate fertilized eggs. Thus these applications biomedical research and clinical medicine create high demand bioactive hLIF. However, production active problematic since eukaryotic cells demonstrate limited expression prokaryotic produce insoluble protein. Here, we have adopted hybrid protein disulfide isomerase design to increase solubility Escherichia coli. Low temperature fused b'a' domain (PDIb'a') increased soluble comparison controls. A simple purification protocol was established includes removal PDIb'a' by cleavage TEV protease. The resulting hLIF, which contains one extra glycine residue at N-terminus, highly pure demonstrated endotoxin levels below 0.05 EU/μg. presence an intramolecular bond identified using mass spectroscopy. This purified effectively maintained murine cell line. developed effective method version E. coli use research.