| 人白介素24单克隆抗体的制备与鉴定 |
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| 引用本文： | 刘世海,李荣华,王东阳,王丽娴,夏海滨.人白介素24单克隆抗体的制备与鉴定[J].现代肿瘤医学,2011,19(9):1701-1705. |
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| 作者姓名： | 刘世海 李荣华 王东阳 王丽娴 夏海滨 |
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| 作者单位： | 陕西师范大学生命科学学院基因治疗研究室,陕西,西安,710062 |
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| 基金项目： | 国家自然科学基金资助项目，陕西师范大学"优秀博士学位论文" |
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| 摘 要： | 目的:制备抗白细胞介素24(Interleukin 24,IL-24)的单克隆抗体并对其生物学特性进行鉴定。方法:应用分子生物学技术,构建含人IL-24编码序列的原核表达载体,表达并纯化了人IL-24融合蛋白。用纯化人IL-24融合蛋白免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,用间接ELISA方法筛选杂交瘤细胞,有限稀释法进行亚克隆,采用免疫印迹及免疫组化染色对抗体的特异性进行了鉴定。结果:获得3株能稳定分泌抗IL-24单抗的杂交瘤细胞,分别命名为1G2,3F4,4A6。通过免疫印迹及免疫组化染色检测显示,这3株单抗均能够与IL-24特异性结合。结论:成功制备抗IL-24单克隆抗体,为进一步探索IL-24在抗肿瘤中的作用机制奠定了基础。
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| 关 键 词： | 白细胞介素24 原核表达 杂交瘤 单克隆抗体 |
Preparation and identification of monoclonal antibodies against interleukin 24 |
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| LIU Shihai,LI Ronghua,WANG Dongyang,WANG Lixian,XIA Haibin.Preparation and identification of monoclonal antibodies against interleukin 24[J].Journal of Modern Oncology,2011,19(9):1701-1705. |
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| Authors: | LIU Shihai LI Ronghua WANG Dongyang WANG Lixian XIA Haibin |
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| Institution: | LIU Shihai,LI Ronghua,WANG Dongyang,WANG Lixian,XIA HaibinLaboratory of Gene Therapy,College of Life Sciences,Shaanxi Normal University,Xi'an 710062,China. |
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| Abstract: | Objective: To prepare and identify the monoclonal antibody against interleukin 24(IL-24).Methods: Prokaryotic expressing vector containing human IL-24 gene was constructed by molecular biology techniques.Then human IL-24 fusion protein was expressed and purified.Splenocytes from the Balb/c mice immunized by human IL-24 fusion protein were fused with myeloma cells SP2/0 for producing hybridoma.Hybridoma cell line secreting antibodies against human IL-24 was determined by indirect ELISA and limiting dilution ... |
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| Keywords: | interleukin 24 prokaryotic expression hybridoma monoclonal antibody |
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