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| 抗人生长转化因子15单克隆抗体制备及鉴定 | |
| 引用本文： | 詹峰,曾晓燕,张晓,周顺,徐言,张建平,焦永军.抗人生长转化因子15单克隆抗体制备及鉴定[J].细胞与分子免疫学杂志,2011,27(5):539-541,544. |
| 作者姓名： | 詹峰 曾晓燕 张晓 周顺 徐言 张建平 焦永军 |
| 作者单位： | 1. 南京医科大学第二附属医院普通外科,江苏,南京,210011 2. 卫生部肠道病原微生物重点实验室江苏省疾病预防控制中心病原微生物研究所,江苏,南京,210009 3. 南京医科大学卫生部抗体技术重点实验室,江苏,南京,210029 |
| 基金项目： | 卫生部科学研究基金资助项目，南京医科大学肿瘤中心招标课题 |
| 摘 要： | 目的:制备抗人生长转化因子15(GDF15)单克隆抗体(mAb)并鉴定其特性。方法:构建GDF15原核表达载体pGEX-4T-2-gdf15,诱导表达并纯化GST-GDF15融合蛋白。以该融合蛋白免疫BALB/c小鼠,取小鼠的脾细胞与同系小鼠Sp2/0骨髓瘤细胞常规融合,依次进行阳性杂交瘤细胞株的筛选及亚克隆,最终获得稳定分泌抗GDF15 mAb杂交瘤细胞株。以间接ELISA法检测抗体效价;Western blot鉴定抗体特异性;免疫共沉淀法初步检测在肝癌患者血清中GDF15表达水平。结果:获得1株稳定分泌抗人GDF15 mAb杂交瘤细胞株,命名为9G3;抗体免疫球蛋白亚类为IgG1,轻链为κ型;免疫共沉淀及质谱分析证实抗GDF15 mAb9G3能与肝癌患者血清中GDF15蛋白特异性结合并且GDF15水平在肝癌患者血清中较正常人显著升高。结论:成功制备了抗人GDF15mAb,为后期肝癌早期诊断试剂盒的开发奠定了良好的基础。 |
| 关 键 词： | GDF15 单克隆抗体 HCC |
Preparation and identification of monoclonal antibody against human growth differentiation factor 15 |
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| ZHAN Feng,ZENG Xiao-yan,ZHANG Xiao,ZHOU Shun,XU Yan,ZHANG Jian-ping,JIAO Yong-jun.Preparation and identification of monoclonal antibody against human growth differentiation factor 15[J].Journal of Cellular and Molecular Immunology,2011,27(5):539-541,544. | |
| Authors: | ZHAN Feng ZENG Xiao-yan ZHANG Xiao ZHOU Shun XU Yan ZHANG Jian-ping JIAO Yong-jun |
| Institution: | ZHAN Feng1,ZENG Xiao-yan2,ZHANG Xiao3,ZHOU Shun1,XU Yan1,ZHANG Jian-ping1,JIAO Yong-jun2 1Department of General Surgery,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210011,2Key Laboratory of Enteric Pathogenic Microbiology,Ministry of Health,Institute of Pathogenic Microbiology,Jiangsu Provincial Center for Disease Prevention and Control,Nanjing 210009,3Key Laboratory of Antibody Technique of Ministry of Health,Nanjing Medical University,Nanjing 210029,China |
| Abstract: | AIM: To prepare the monoclonal antibody(mAb) against human growth differentiation factor 15(GDF15).METHODS: The expression vector pGEX-4T-2-gdf15 was constructed and transformed into E.coli Top10F' for expression.Then the purified fusion protein was used to immunize the BALB/c mice.The mouse myeloma cells(Sp2/0) were fused with spleen cells from the BALB/c immunized by the purified protein.Subsequently,limited dilution method was used three times to screen hybridoma cell lines.The titer of the mAb was detec... |
| Keywords: | GDF15 HCC |
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