| 苦马豆素单克隆抗体的制备与鉴定 |
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| 引用本文： | 贾琦珍,陈根元,王连群,亚森&#,麦麦提,王帅.苦马豆素单克隆抗体的制备与鉴定[J].中国畜牧兽医,2017,44(7):2155-2159. |
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| 作者姓名： | 贾琦珍 陈根元 王连群 亚森&# 麦麦提 王帅 |
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| 作者单位： | 塔里木大学动物科学学院, 新疆生产建设兵团塔里木畜牧科技重点实验室, 阿拉尔 843300 |
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| 基金项目： | 国家自然科学基金（31460678）；国家星火计划项目（2015GA891015）；兵团塔里木畜牧科技重点实验室开放课题（HS201207） |
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| 摘 要： | 本试验采用苦马豆素（swainsonine，SW）人工抗原SW-OVA免疫接种BALB/c小鼠，通过杂交瘤技术建立了1株能稳定分泌SW单克隆抗体的杂交瘤细胞1F10，杂交瘤细胞染色体数目为45~50对。经间接ELISA检测，该株细胞培养上清中抗体效价为1：25 600，诱生腹水效价为1：80 000。单克隆抗体的亚型为IgG1，亲和力解离常数为1.14×1010，腹水抗体纯化后纯度可达98%，回收率80%，SDS-PAGE凝胶电泳可见单克隆抗体的轻链、重链分子质量分别为25和50 ku，Western blotting检测抗体能特异性的识别SW。其线性检测范围为4~128 μg/mL（R2=0.9969），与BSA、明胶、多聚赖氨酸、α-甘露糖苷等无交叉反应。本试验制备的SW单克隆抗体为免疫学检测SW和免疫学防制家畜疯草中毒奠定基础。
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| 关 键 词： | 苦马豆素 单克隆抗体 杂交瘤细胞 间接ElISA |
| 收稿时间： | 2017-01-18 |
Preparation and Identification of Monoclonal Antibody Against Swainsonine |
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| JIA Qi-zhen,CHEN Gen-yuan,WANG Lian-qun,YASEN&#,Maimaiti,WANG Shuai.Preparation and Identification of Monoclonal Antibody Against Swainsonine[J].China Animal Husbandry & Veterinary Medicine,2017,44(7):2155-2159. |
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| Authors: | JIA Qi-zhen CHEN Gen-yuan WANG Lian-qun YASEN&# Maimaiti WANG Shuai |
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| Institution: | Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production & Construction Corps, College of Animal Science, Tarim Unversity, Alar 843300, China |
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| Abstract: | In this test,BALB/c mice were immunized by SW-OVA for the preparation of monoclonal antibody against swainsonine (SW). Hybridoma cells that could secrete specific antibody against SW were prepared by hybridoma technique,and then the strain that secreted monoclonal antibody against SW designated 1F10 was prepared,and the chromosome average number of 1F10 cell was 45 to 50 couples. The ELISA titers of cell supernatant were 1:25 600, and that of ascites were 1:80 000.The subclasses of monoclonal antibody was IgG1,and the affinity constant was 1.14×1010,the purity of ascites antibodies was up to 98%,and the recovery rate was 80%.The result of sodium dodecyl sulfate polyacrylamide gel electropheresis proved that purified antibody had been obtained that the molecular weight of H-chain and L-chain of antibody was about 50 and 25 ku. The monoclonal antibody against SW specifically bound to SW determined by Western blotting. The linear range was 4 to 128 μg/mL (R2=0.9969) and no cross-reactivity was detected with BSA,gelatin,polylysine,me-Gal,etc. The result laid the foundation for immunodetection on SW and immunological prevention of animal toxic disease. |
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| Keywords: | swainsonine monoclonal antibody hybridoma cell indirect ElISA |
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