| 抗猪瘟病毒单克隆抗体的制备及特性鉴定 |
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| 引用本文： | 王爱华,许可军,王金良,魏凤,孙全文.抗猪瘟病毒单克隆抗体的制备及特性鉴定[J].中国畜牧兽医,2014,41(11):97-100. |
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| 作者姓名： | 王爱华 许可军 王金良 魏凤 孙全文 |
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| 作者单位： | 1. 河北北方学院, 河北张家口 075000;2. 滨州市农村科技信息化管理中心, 山东滨州 256600;3. 山东省滨州畜牧兽医研究院, 山东滨州 256600 |
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| 基金项目： | 河北北方学院重大项目(ZD201307);山东省自然科学基金项目(ZR2010CQ012);山东省大型科学仪器设备升级改造技术研究专项(2013SJGZ18)。 |
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| 摘 要： | 将纯化后的猪瘟病毒免疫4只SPF级BALB/c小鼠,无菌取脾细胞与SP2/0骨髓瘤细胞融合,经半固体培养基克隆化和间接ELISA筛选,最终获得了4株稳定分泌抗猪瘟病毒单克隆抗体的杂交瘤细胞株。ELISA结果显示4株单克隆抗体效价均较高,并与其他病毒及PK-15细胞培养物无交叉反应。Western blotting结果证实3株单克隆抗体可特异性识别猪瘟病毒。间接免疫荧光试验可在细胞膜内观察到特异性绿色荧光,表明3株单克隆抗体与猪瘟病毒具有良好的反应性。该研究为猪瘟病毒新型诊断试剂开发奠定了基础。
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| 关 键 词： | 猪瘟病毒 单克隆抗体 制备 鉴定 |
| 收稿时间： | 2014-06-03 |
Preparation and Characteristic Identification of Monoclonal Antibodies against Classical Swine Fever Virus |
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| WANG Ai-hua,XU Ke-jun,WANG Jin-liang,WEI Feng,SUN Quan-wen.Preparation and Characteristic Identification of Monoclonal Antibodies against Classical Swine Fever Virus[J].China Animal Husbandry & Veterinary Medicine,2014,41(11):97-100. |
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| Authors: | WANG Ai-hua XU Ke-jun WANG Jin-liang WEI Feng SUN Quan-wen |
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| Institution: | 1. Hebei North University, Zhangjiakou 075000, China;2. Binzhou Rural Science and Technology Information Management Center, Binhzou 256600, China;3. Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, China |
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| Abstract: | 4 SPF BALB/c mice were immunized with purified classical swine fever virus (CSFV) as the antigen and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA and semi-solid agar cloning, 4 hybridoma cells secreting anti-CSFV monoclonal antibodies were obtained. ELISA results showed that all of these monoclonal antibodies with high antibody titer, which could specifically react to CSFV but not cross react to other pig viruses and PK-15 cell cultures. Western blotting indicated that 3 strains of monoclonal antibodies reacted with CSFV specially. Specific green fluorescence was observed in the cell membrane by IFA. This study layed a foundation for the development of new diagnostic reagents for CSFV. |
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| Keywords: | classical swine fever virus (CSFV) monoclonal antibody preparation identification |
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