| Pfs25蛋白单克隆抗体的制备及双抗体夹心ELISA的建立 |
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| 引用本文： | 李萍,马亚茹,陆俭,雷清,陈勇,蒋琳. Pfs25蛋白单克隆抗体的制备及双抗体夹心ELISA的建立[J]. 细胞与分子免疫学杂志, 2011, 27(12): 1330-1334 |
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| 作者姓名： | 李萍 马亚茹 陆俭 雷清 陈勇 蒋琳 |
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| 作者单位： | 国药集团兰州生物制品研究所,甘肃兰州,730046 |
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| 摘 要： | 目的:制备恶性疟原虫子孢子囊表面膜蛋白Pfs25的单克隆抗体( mAb),建立检测Pfs25蛋白的双抗体夹心ELISA方法.方法:纯化毕赤酵母表达的重组Pfs25蛋白,并免疫BALB/c小鼠,采用骨髓瘤细胞Sp2/0与免疫BALB/c鼠脾细胞杂交的细胞融合技术,通过间接ELISA检测获得分泌抗Pfs25抗体的阳性杂交瘤细胞株,通过免疫F1鼠诱生腹水,纯化腹水,并进行mAb的各项生物学鉴定.辣根过氧化物酶(HRP)标记纯化后的抗体,以4B7为包被抗体,1B4为酶标抗体,建立了双抗体夹心ELISA法.结果:获得3株抗Pfs25的杂交瘤细胞株,其中2株有良好的稳定性和特异性.并建立了双抗体夹心ELISA检测法,检测有效范围在0.07～1 mg/mL,其检测灵敏度为41.6 ng/mL.结论:成功制备抗Pfs25蛋白的单克隆抗体,并建立了一种可用于Pfs25蛋白检测的双抗体夹心ELISA法,为Pfs25蛋白制备传播阻断型疟疾疫苗奠定了基础.
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| 关 键 词： | Pfs25蛋白 单克隆抗体 双抗体夹心ELISA 传播阻断型疟疾疫苗 |
Preparation of monoclonal antibody against Pfs25 protein and establishment of double antibody sandwich ELISA method |
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| LI Ping , MA Ya-ru , LU Jian , LEI Qing , CHEN Yong , JIANG Lin. Preparation of monoclonal antibody against Pfs25 protein and establishment of double antibody sandwich ELISA method[J]. Chinese journal of cellular and molecular immunology, 2011, 27(12): 1330-1334 |
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| Authors: | LI Ping MA Ya-ru LU Jian LEI Qing CHEN Yong JIANG Lin |
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| Affiliation: | LI Ping,MA Ya-ru,LU Jian,LEI Qing,CHEN Yong,JIANG Lin Lanzhou Institute of Biological Products,Lanzhou 730046,China |
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| Abstract: | AIM: To prepare the monoclonal antibody(mAb)against Pfs25 protein of Plasmodium falciparum,and establish the method of sandwich ELISA for detecting the Pfs25 protein.METHODS: Pfs25 protein the recombinant expressed by Pichia pastoris was purified.The purified Pfs25 protein as the antigen was used to immune the BALB/c mice,The secreting specific mAb positive cell strains,which were prepared by hybridizing the Sp2/0 myeloma cell and the spleen cell from immunized mice,were detected by indirect ELISA method.Th... |
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| Keywords: | Pfs25 protein monoclonal antibody double antibodies sandwich ELISA transmission-blocking malaria vaccine |
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