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苦马豆素单克隆抗体的制备与鉴定
  • ISSN号:1671-7236
  • 期刊名称:《中国畜牧兽医》
  • 时间:0
  • 分类:R392[医药卫生—免疫学;医药卫生—基础医学]
  • 作者机构: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
  • 相关基金: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)
中文摘要:

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50ku,Western blotting检测抗体能特异性的识别SW。其线性检测范围为4~128μg/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。

英文摘要:

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 50couples.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 about50 and 25ku.The monoclonal antibody against SW specifically bound to SW determined by Western blotting.The linear range was 4to 128μg/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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期刊信息
  • 《中国畜牧兽医》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院北京畜牧兽医研究所
  • 主编:李琍
  • 地址:中国农业科学院北京畜牧兽医研究所
  • 邮编:100193
  • 邮箱:zgxmsy@caas.cn
  • 电话:010-62816020 62811226 62810371
  • 国际标准刊号:ISSN:1671-7236
  • 国内统一刊号:ISSN:11-4843/S
  • 邮发代号:2-215
  • 获奖情况:
  • 1996年被农业部评为“95中国出版成就展”参展期刊...,1998年在首届全国畜牧兽医优秀科技期刊评比中总分...,2002年被中国期刊协会农业期刊分会和中国农学会评...
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:22619