| 癌-睾丸抗原OY-TES-1多克隆抗体的制备及鉴定 |
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| 引用本文： | 范蓉,肖绍文,黄巍,罗彬,李颀,徐丽湘,谢小薰.癌-睾丸抗原OY-TES-1多克隆抗体的制备及鉴定[J].粉末涂料与涂装,2009,22(8). |
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| 作者姓名： | 范蓉 肖绍文 黄巍 罗彬 李颀 徐丽湘 谢小薰 |
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| 作者单位： | 范蓉(广西中医学院组胚教研室,南宁,530001);肖绍文,罗彬,李颀,徐丽湘,谢小薰(广西医科大学组胚教研室,南宁,530021);黄巍(广西中医学院第一附属跃院妇产科,南宁,530023) |
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| 基金项目： | 国家自然科学基金，国家自然科学基金，广西自治区科学基金 |
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| 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。
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| 关 键 词： | 癌-睾丸抗原 OY-TES-1 融合蛋白 多克隆抗体 |
Preparation and Identification of Polyclonal Antibody against Cancer-Testis Antigen OY-TES-1 |
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| Abstract: | 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.coli,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 immunohisto-chemical 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. |
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| Keywords: | OY-TES-1 |
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