摘要
目的制备针对青霉素结合蛋白(PBP2a)的抗血清,比较不同免疫方案的免疫效果。方法以基因工程重组PBP2a转肽酶区蛋白作为免疫原,采用多种注射途径(足垫、腹腔、足垫+腹腔、足垫+皮下)免疫BALB/C小鼠,ELISA和Western-blot法检测所制备的抗血清效价和特异性。结果足垫快速免疫组、腹腔免疫组、足垫和腹腔免疫组、足垫加皮下免疫组的效价依次为1∶12800、1∶25600、1∶51200、1∶102400,Western-blot显示所制备的抗血清能有效识别原核表达及耐甲氧西林的金黄色葡萄球菌(MRSA)临床分离株中的PBP2a。结论重组PBP2a蛋白免疫BALB/c小鼠能有效刺激特异性抗体的产生,足垫加皮下多点免疫法是一种值得推崇的免疫方案。
Objective To prepare mouse polyclonal antibody against penicillin binding protein 2a （PBP2a） generated by different immunization protocols. Method BALB/C mice were immunized with the recombinant transpeptidase domain of PBP2a by different injection routes （foot pad, intraperitioneal, foot pad ＋subcutaneous and foot pad ＋intraperitioneal）.The titer and the specificity of the anti-PBP2a antibodies were detected by ELISA and Western-blot. Result The titers of anti-PBP2a antibodies of the mice immunized by foot pad, intraperitioneal, foot pad＋subcutaneous and foot pad＋intraperitioneal were 1：12800, 1：25600, 1：51200, and 1：102400, respectively. Western-blot analysis showed that the polyclonal anti-PBP2a antibodies could detect both prokaryotic expressed PBP2a, and clinical isolated MRSA-PBP2a. Conclusion Polyclonal anti-PBP2a antibodies with high specificity were prepared by using recombinant PBP2a as immunogen. Foot pad ＋intraperitiuneal immunization produce the highest titer of specific antibodies.
出处
《热带医学杂志》
CAS
2007年第2期145-147,共3页
Journal of Tropical Medicine
基金
广州市医药卫生科技项目(No.2005-YB-135)
广州医学院项目(No.03-K-32)
