摘要
目的制备癌-睾丸抗原OY-TES-1多克隆抗体,并进行鉴定。方法以大肠杆菌表达的MBP/OY-TES-1融合蛋白作为抗原,免疫新西兰白兔,制备抗血清。经蛋白A凝胶纯化后,采用ELISA法检测抗血清的效价,Western blot和免疫组化法检测多克隆抗体的特异性。结果制备的抗血清经ELISA检测,效价不低于1∶128000,纯化后,其效价不低于1∶8000。Western blot显示,抗OY-TES-1多抗可与MBP/OY-TES-1融合蛋白、MBP、OY-TES-1重组蛋白和睾丸组织总蛋白特异性结合;免疫组化检测显示,精子头部呈现阳性反应。结论已成功制备了效价高、特异性强的癌-睾丸抗原OY-TES-1多克隆抗体,为深入研究OY-TES-1的生物学功能提供了工具。
Objective To prepare and identify the polyclonal antibody against cancer-testis antigen OY-TES-1. Methods New Zealand rabbits were immunized with MBP/OY-TES-1 fusion protein expressed in E. colt, and the obtained antisera were purified by protein A gel chromatography, then determined for titer by ELISA and tested for specificity by Western blot and immunohistochemical assay. Results The ELISA titers of prepared antisera were not less than 1 ： 128 000 and not less than 1 ： 8 000 before and after purification respectively. Western blot showed specific reactions of the polyclonal antibody with MBP/OY-TES-1 fusion protein, MBP, recombinant OY-TES-1 protein and total protein of testis tissue. Immunohistochemical assay revealed the protein bound to the polyclonal antibody in human spermids. Conclusion The polyclonal antibody against cancer-testis antigen OY-TES-1, with high titer and specificity, was successfully prepared, which provided a tool for further study on the biological function of OY-TES-1.
出处
《中国生物制品学杂志》
CAS
CSCD
2009年第8期804-806,共3页
Chinese Journal of Biologicals
基金
国家自然科学基金(30360026)
国家自然科学基金(30760055)
广西科学基金(桂科基0639040)
