Purification of the LukF-PV subunit of Staphylococcus aureus Panton-Valentine leukocidin expressed in Escherichia coli and generation of antiserum in mice

Abstract

LukF-PV is one of the two subunits of the virulence factor Panton-Valentine leukocidin (PVL) produced by Staphylococcus aureus. The LukF-PV subunit can bind with the LukS-PV subunit in a 4:4 ratio to construct an octameric structure that creates pores in the leukocyte cell membrane, leading to leukocyte destruction. This study focused on purifying the LukF-PV protein fused with a 6xHis-Tag (6xHis-LukF-PV) and raising antiserum in mice. The expression of 6xHis-LukF-PV in Escherichia coli BL21(DE3) was achieved under culture conditions with appropriate temperature, inducer and time after induction, resulting in an expression level of approximately 30% of total intracellular protein. Using affinity chromatography, 6xHis-LukF-PV was purified to 95% purity, achieving a concentration of 2.45 mg/ml and a yield of 6.75 mg/l in LB medium. Antisera were generated in mice, having antiserum with a titer of 1/160,000. The results provide information and primary materials for future preventative or treatment studies against S. aureus.