CAJ | 학술논문

目的：制备抗小鼠ZP2（mZP2）抗体并研究抗血清的免疫活性。方法利用Rosetta宿主菌诱导表达mZP2重组蛋白，电泳分离重组ZP2蛋白，切胶制备免疫抗原；利用乳化抗原主动免疫新西兰雄兔制抗血清；通过ELISA测定抗体应答水平；以免疫组化测定抗血清特异性；通过精卵结合实验检测抗血清对小鼠卵子结合顶体反应后精子的影响。结果 ELISA结果说明用重组蛋白免疫新西兰雄兔诱发产生了抗血清。免疫组化实验证明抗血清与小鼠卵巢透明带发生了特异性反应，精卵结合实验显示抗ZP2抗体能抑制顶体反应后精子与卵子结合。结论重组mZP2蛋白诱发产生了抗ZP2抗体，抗体能抑制顶体反应后的精子与卵子结合。
목적：제비항소서ZP2（mZP2）항체병연구항혈청적면역활성。방법이용Rosetta숙주균유도표체mZP2중조단백，전영분리중조ZP2단백，절효제비면역항원；이용유화항원주동면역신서란웅토제항혈청；통과ELISA측정항체응답수평；이면역조화측정항혈청특이성；통과정란결합실험검측항혈청대소서란자결합정체반응후정자적영향。결과 ELISA결과설명용중조단백면역신서란웅토유발산생료항혈청。면역조화실험증명항혈청여소서란소투명대발생료특이성반응，정란결합실험현시항ZP2항체능억제정체반응후정자여란자결합。결론중조mZP2단백유발산생료항ZP2항체，항체능억제정체반응후적정자여란자결합。
Objective To prepare rabbit anti- mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity. Methods Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay. Results ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%. Conclusion We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.