| 抗人Cu／Zn SOD单克隆抗体的制备与鉴定 |
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| 引用本文： | 蒋小梅,廖婷婷,章崇杰,张晨阳,魏大鹏. 抗人Cu／Zn SOD单克隆抗体的制备与鉴定[J]. 西部医学, 2010, 22(4): 603-606,609 |
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| 作者姓名： | 蒋小梅 廖婷婷 章崇杰 张晨阳 魏大鹏 |
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| 作者单位： | 四川大学基础医学与法医学院免疫学教研室,四川,成都,610041 |
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| 摘 要： | 目的制备抗人Cu／Zn SOD单克隆抗体并对其生物学特性进行鉴定。方法以人Cu／Zn SOD作为免疫原免疫Balb／c小鼠，取其脾细胞与SP2／0细胞融合，经多次筛选及克隆化，建立可稳定分泌抗人Cu／Zn SOD单克隆抗体的杂交瘤细胞株。用ELISA、斑点ELISA、Western Blot、免疫组化鉴定单克隆抗体的特性，并测定其效价、亚型及相对亲和力。结果筛选到一株可稳定分泌抗人Cu／Zn SOD单克隆抗体的杂交瘤细胞株4H10H4B12，亚型鉴定重链为IgG2a，轻链为kappa链。ELISA法测定腹水效价为1：256，000，抗体相对亲和力为1．101×10^9mol／L。斑点ELISA、Western Blot显示抗体能特异识别免疫原、人肝细胞株HL-7702和人红细胞中的天然Cu／Zn SOD；不与鼠Cu／Zn SOD和人Mn SOD发生交叉反应。免疫组化进一步显示了人Cu／Zn SOD均匀分布于细胞的胞浆中。结论成功制备了抗人Cu／Zn SOD单克隆抗体，为进一步研究其与腹主动脉瘤的关系奠定了基础。
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| 关 键 词： | 人Cu／Zn SOD 单克隆抗体 腹主动脉瘤 |
Preparation and identification of monoclonal antibody against human Cu/Zn SOD |
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| JIANG Xiao-mei,LIAO Ting-ting,ZHANG Chen-yang,et al. Preparation and identification of monoclonal antibody against human Cu/Zn SOD[J]. , 2010, 22(4): 603-606,609 |
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| Authors: | JIANG Xiao-mei LIAO Ting-ting ZHANG Chen-yang et al |
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| Affiliation: | JIANG Xiao-mei,LIAO Ting-ting,ZHANG Chen-yang,et al(Department of Immunology,School of Preclinical , Forensic Medicine,Sichuan University,Chengdu 610041,China) |
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| Abstract: | Objective To prepare and identify the monoclonal antibody(MAb) against human Cu/Zn SOD.Methods BALB/C mice were immunized with immunogen human Cu/Zn SOD.The spleen cells of the immunized mice were isolated and fused with SP2/0 cells.After several rounds of detecting and cloning,a hybridoma cell line secreting anti-human Cu/Zn SOD MAb was obtained.Its specificity was evaluated with indirect-ELISA,spot-ELISA,Western blot and immunohistochemistry.The titer,immunoglobulin subtype and affinity of the MAb were me... |
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| Keywords: | SOD |
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