| 抗重金属Cr3+单克隆抗体的制备及其应用 |
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| 引用本文： | 颜露,唐勇,向军俭,翟一凡,杨红宇.抗重金属Cr3+单克隆抗体的制备及其应用[J].细胞与分子免疫学杂志,2011,27(4):422-424. |
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| 作者姓名： | 颜露 唐勇 向军俭 翟一凡 杨红宇 |
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| 作者单位： | 暨南大学分子免疫与抗体工程中心,广东广州,510632 |
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| 基金项目： | 国家“863”高技术发展计划目标导向项目(2008AA10Z427) |
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| 摘 要： | 目的:制备重金属Cr3+的单克隆抗体(mAb)并建立间接竞争ELISA法,用于Cr3+的检测。方法:用合成的重金属铬-螯合剂-BSA抗原免疫BALB/c小鼠,通过细胞融合、克隆化和ELISA等技术建立杂交瘤细胞株;腹水型mAb经硫酸铵沉淀纯化后建立间接竞争ELISA法并与9种金属进行交叉实验;水样加标后,进行间接竞争ELISA法与ICP-AES法检测Cr3+的比较,计算两种方法符合率。结果:获得3株稳定分泌抗重金属Cr3+mAb的杂交瘤细胞株,其中5G12C5株mAb间接ELISA效价>1∶1×105,间接竞争ELISA法检测Cr3+的范围为0.783~50μg/L,最低检测限为1μg/L,与Fe3+存在<10%交叉,而与其他金属无交叉反应;当水样中加标量>1μg/L时,间接竞争ELISA法与ICP-AES法检测Cr3+的符合率达96.95%。结论:获得效价高、特异性强的重金属Cr3+mAb,初步建立了检测Cr3+的间接竞争ELISA法,该方法检测的灵敏度可达到国家地下Ⅰ类水的检测标准(0.005 mg/L)。
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| 关 键 词： | 重金属 Cr3+ 细胞融合 单克隆抗体 ELISA |
Preparation and application of monoclonal antibodies against Heavy Metal Chromium |
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| YAN Lu,TANG Yong,XIANG Jun-jian,ZHAI Yi-fan,YANG Hong-yu.Preparation and application of monoclonal antibodies against Heavy Metal Chromium[J].Journal of Cellular and Molecular Immunology,2011,27(4):422-424. |
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| Authors: | YAN Lu TANG Yong XIANG Jun-jian ZHAI Yi-fan YANG Hong-yu |
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| Institution: | YAN Lu,TANG Yong,XIANG Jun-jian,ZHAI Yi-fan,YANG Hong-yu Antibody Engineering Center,Jinan University,Guangzhou 510632,China |
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| Abstract: | AIM:To produce the monoclonal antibodies(mAb) against Heavy Metal Chromium,and develop a competitive inhibition enzyme-linked immunosorbent assay(ciELISA) for the detection of Chromium.METHODS: Chromium ions was chelated with bifunctional chelating agent Isothio-cyanobenzyl-EDTA firstly and then conjugated with bovin serum alburmin(BSA) and ovalbmin(OVA) respectively to complete antigens.Four female BALB/c mice were immunized with Cr-iEDTA-BSA,the Hybridoma lines secreting monoclonal antibodies against Chro... |
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| Keywords: | heavy metal chromium cell fusion mAb ELISA |
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