| 抗鹅细小病毒VP3蛋白单克隆抗体的制备及鉴定 |
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| 引用本文： | 郭鹭,荆志强,李俚,鞠环宇,马波,王君伟.抗鹅细小病毒VP3蛋白单克隆抗体的制备及鉴定[J].中国兽医科学,2010,40(2). |
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| 作者姓名： | 郭鹭 荆志强 李俚 鞠环宇 马波 王君伟 |
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| 作者单位： | 东北农业大学,动物医学学院,黑龙江,哈尔滨,150030 |
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| 基金项目： | 黑龙江省教育厅重大项目，黑龙江省攻关项目 |
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| 摘 要： | 利用已构建好的pET-30a重组菌E.coli Rosetta(DE3)表达鹅细小病毒(GPV)VP3重组蛋白,经Ni-NTA亲和层析纯化后作为免疫原,经腹膜腔接种4～6周龄BALB/c小鼠3次,末次免疫后第3d取小鼠脾细胞与SP2/0骨髓瘤细胞进行融合。用建立的间接ELISA方法进行筛选,经4次亚克隆,最终获得4株能稳定分泌抗GPVVP3单克隆抗体的杂交瘤细胞株,分别命名为1F1、2A9、3B11和4A2。亚类鉴定结果显示,单抗1F1为IgG2b亚型,其余3株单抗为IgG1亚型,轻链均为κ链。经间接ELISA检测,单抗1F1、2A9、3B11和4A2的腹水效价分别为1:819200、1:1638400、1:409600和1:819200。Western-blot分析表明,获得的4株单克隆抗体均能特异性识别GPVVP3重组蛋白;间接免疫荧光试验证明,这4株单抗能够与天然GPV结合。
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| 关 键 词： | 鹅细小病毒 VP3重组蛋白 单克隆抗体 特性鉴定 |
Preparation and identification of monoclonal antibodies against VP3 protein of goose parvovirus |
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| GUO Lu,JING Zhi-qiang,LI Li,JU Huan-yu,MA Bo,WANG Jun-wei.Preparation and identification of monoclonal antibodies against VP3 protein of goose parvovirus[J].Veterinary Science in China,2010,40(2). |
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| Authors: | GUO Lu JING Zhi-qiang LI Li JU Huan-yu MA Bo WANG Jun-wei |
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| Institution: | GUO Lu,JING Zhi-qiang,LI Li,JU Huan-yu,MA Bo,WANG Jun-wei(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The goose parvovirus(GPV) VP3 protein expressed in the recombinant E.coli Rosetta(DE3) was used as immunogen after purification by Ni-NTA.The 4 to 6 weeks-old BALB/c mice were intraperitoneally immunized with the VP3 protein for three times,then myeloma cells SP2/0 were fused with the spleen cells of the immunized BALB/c mice on day 3 post-last-immunization.Four hybridoma cell lines against the VP3 protein were obtained by screening with the indirect ELISA,named 1F1,2A9,3B11 and 4A2,respectively.The first m... |
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| Keywords: | goose parvovirus recombinant VP3 protein monoclonal antibody specific identification |
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