摘要
目的制备抗小鼠ZP2(m ZP2)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m ZP2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m ZP2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。
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.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2015年第4期522-525,548,共5页
Journal of Southern Medical University
基金
国家自然科学基金重点项目(21307017)
河北省2014年度医学科学研究重点课题(ZD20140047)~~
