重组PV-杀白细胞素LukF-PV蛋白多克隆抗体的制备及鉴定

Production and Verification of Polyclonal Antibodies with Rocombinant LukF-PV

目的制备重组金黄色葡萄球菌PV-杀白细胞素LukF-PV蛋白多克隆抗体并鉴定。方法纯化获得重组蛋白LukF-PV,免疫新西兰白兔,收集多抗血清,ELISA法测定抗体滴度,Western blotting法鉴定免疫活性。结果成功免疫新西兰白兔获得多抗血清,ELISA测定其效价为1∶103,Western blotting鉴定其能识别重组蛋白和金葡菌PVL蛋白。结论成功制备出重组金黄色葡萄球菌PV-杀白细胞素LukF-PV蛋白多克隆抗体,并进行鉴定,为建立产杀白细胞素的金黄色葡萄球菌的快速、廉价的免疫学检测方法奠定基础。

Objective The polyclonal antibodies obtained through the immunization of New Zealand white rabbits with recombinant LukF-PV.Methods Fusion expression was induced by IPTG and purified through affinity chromatography with NiSO4. New Zealand white rabbits were immunized with recombinant LukF-PV （r LukF-PV） and sera of the immunized rabbits were collected 10 days after the second enhanced immunization. The polyclonal antibodies thus obtained were verified with rLukF-PV antigen and PVL by western blotting and the titers of antibodies were determined by ELISA.Results The polyclonal antibodies obtained through the immunization of New Zealand white rabbits with rLukF-PV showed a serum antibody titer of 1∶103. Conclusion The polyclonal antibodies were obtained successfully. The results in the present study may provide a foundation for the futher studies on the diagnosis of PVL.