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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (11): 31-34.

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

抗犬I型腺病毒单克隆抗体的制备及生物学特性鉴定

袁宝 刘颖 蓝田丰 艾纯旭 陈健 任文陟 李晓叶 刘殿峰   

  • 收稿日期:2010-12-14 修回日期:2010-12-31 出版日期:2011-05-15 发布日期:2011-05-15
  • 基金资助:

    吉林省科技平台建设项目“吉林省实验动物质量检测中心平台建设”（20071138）

Preparation of Monoclonal Antibodies against Canine Adenovirus TypeⅠ and Their Identification of Biological Characters

  • Received:2010-12-14 Revised:2010-12-31 Online:2011-05-15 Published:2011-05-15

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摘要：

摘要：目的　制备抗犬I型腺病毒单克隆抗体。方法　犬I型腺病毒(CAV-I)细胞培养液经饱和硫酸铵沉淀，差速离心浓缩，氯化铯密度梯度离心纯化后免疫BALB/c小鼠，三免后效价过1：10000即可取脾细胞与SP2/0细胞在聚乙二醇（PEG）作用下融合，通过间接ELISA方法筛选阳性杂交瘤细胞株，有限稀释法亚克隆，制备单克隆抗体，并对制备完成的单克隆抗体进行生物学特性鉴定。结果　获得2株能稳定分泌抗CAV-I的单克隆抗体杂交瘤细胞，命名为C8、E9，经鉴定其亚型分别为IgG1和IgG2a。经ELISA检测，2株单抗的细胞上清液效价为1∶1600~1∶3200，其腹水效价为1∶25600~1∶51200。该单克隆抗体与CDV、FPV、FCV病毒均无交叉反应。结论　成功制备了抗CAV-I单克隆抗体，为进一步建立相关诊断方法奠定了基础。

关键词: 金融市场, 金融市场

Abstract:

Abstract:Objective To prepare and identify the monoclonal antibody against canine adenovirus typeⅠ. Methods cell culture fluid of CAV-I were precipitationed by saturated ammonium sulfate, concentrated differential centrifugation, caesium chloride density gradient centrifugation,BALB/c mice were immunized conventionally with the purified CAV-I. On the third day after the final immunization,spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively,detecting special antibody,ELISA test, and coloning. Two hybridoma cell lines,obtained by culturing in vitro and anabiosis after freezing in a long period which were named C8、E9,could stably secrete McAbs. Results　The subclasses of the two McAbs were IgG2a and IgG1 respectively. Among the two McAbs,the titers of cell cultures ranged from 1∶1600 to 1∶3200 and the titers of ascites fluids ranged from 1∶25600 to 1∶51200. These McAbs were specific to CAV-I,but not reacted with CDV、FPV、FCV.Conclusion McAb against CAV-I has been successfully prepared. It is also a basic for establishing a related diagnoses method.