| 抗HCV HVR1模拟合成肽单克隆抗体的制备及特性鉴定 |
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| 引用本文： | 孙杰,戚扬,迟淑萍,由莉越,李瑞生,刘佳,朱雷,程云.抗HCV HVR1模拟合成肽单克隆抗体的制备及特性鉴定[J].细胞与分子免疫学杂志,2011,27(5):542-544. |
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| 作者姓名： | 孙杰 戚扬 迟淑萍 由莉越 李瑞生 刘佳 朱雷 程云 |
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| 作者单位： | 解放军302医院传染病研究所免疫室,北京,100039 |
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| 摘 要： | 目的:制备抗-HCV高变区1(HVR1)合成肽单克隆抗体(mAb),并对其特性进行鉴定。方法:以固相合成的HCV HVR1合成肽与牛血清白蛋白(BSA)偶联后,免疫8周龄的BALB/c小鼠。采用杂交瘤技术,取免疫小鼠的脾细胞与骨髓瘤细胞融合,制备抗-HCV HVR1 mAb。经HAT、HT选择培养及有限稀释法进行克隆化后,用相关的试剂盒鉴定小鼠mAb的Ig亚类;经中和抑制后用ELISA法检测mAb的特异性;用间接ELISA法检测mAb腹水的效价及相对亲和常数。结果:获得1株可分泌抗-HCV HVR1合成肽特异性mAb的杂交瘤细胞1A9G9F11。该株mAb的Ig亚类为IgG3;腹水效价为3.125×10-5;相对亲和常数为1.0×106L/mol。该株mAb与HBsAg、HBeAg、HBcAg、HAAg、牛血清白蛋白、酪蛋白和胸腺5肽均无交叉反应,与HCV HVR1合成肽-牛血清白蛋白出现特异性反应。结论:成功地建立了1株可分泌抗-HCV HVR1 mAb的杂交瘤细胞1A9G9F11,为进一步的实验研究提供了重要的制剂。
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| 关 键 词： | HCVHVR1 合成肽 单克隆抗体 特性鉴定 |
Preparation and characterization of the anti-HCV HVR1 synthetic peptide monoclonal antibody |
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| SUN Jie,QI Yang,CHI Shu-ping,YOU Li-yue,LI Rui-sheng,LIU Jia,ZHU Lei,CHENG Yun.Preparation and characterization of the anti-HCV HVR1 synthetic peptide monoclonal antibody[J].Journal of Cellular and Molecular Immunology,2011,27(5):542-544. |
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| Authors: | SUN Jie QI Yang CHI Shu-ping YOU Li-yue LI Rui-sheng LIU Jia ZHU Lei CHENG Yun |
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| Institution: | SUN Jie,QI Yang,CHI Shu-ping,YOU Li-yue,LI Rui-sheng,LIU Jia,ZHU Lei,CHENG Yun Department of Immunology,Institute of Infections Diseases,302nd Hospital of Chinese PLA,Beijing 100039,China |
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| Abstract: | AIM: To prepare monoclonal antibody(mAb) against HCV HVR1 synthetic peptide and characterize its property.METHODS: 8 week-old female BALB/c mice were immunized with HCV HVR1 synthetic peptide which was linked to bovine serum albumin(BSA) as immunogen.The splenocytes of immunized mice were fused with Sp2/0 cells by hybridoma technique.Monoclonal hybridoma cell line was obtained by limiting dilution,HAT and HT-selective culture.The specificity of mAb was identified by indirect ELISA,immunohistochemical staini... |
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| Keywords: | HCV HVR1 |
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