一株禽腺病毒4型Fiber2蛋白单克隆抗体的制备

doi:10.14088/j.cnki.issn0439-8114.2022.10.021

湖北农业科学 ›› 2022, Vol. 61 ›› Issue (10): 121-126.doi: 10.14088/j.cnki.issn0439-8114.2022.10.021

一株禽腺病毒4型Fiber2蛋白单克隆抗体的制备

谢明, 李国攀, 胡基雄, 张志翔, 艾笑扬, 陈帝斯, 李杨, 尹俊翔, 荣俊

长江大学生命科学学院,湖北荆州 434020

收稿日期:2022-01-19 出版日期:2022-05-25 发布日期:2022-06-14
通讯作者: 荣俊,男,湖北石首人,教授,博士,研究方向为动物传染病免疫学,（电子信箱）496281344@qq.com。
作者简介:谢明（1997-）,男,湖北潜江人,硕士,研究方向为动物疾病预防,（电话）17721867076（电子信箱）1052149811@qq.com。
基金资助:
青岛易邦公司联合开发横向课题“禽腺病毒亚单位疫苗开发利用”（2016）

Preparation of the monoclonal antibody against the serotype 4 fowl adenovirus fiber2 protein

XIE Ming, LI Guo-pan, HU Ji-xiong, ZHANG Zhi-xiang, AI Xiao-yang, CHEN Di-si, LI Yang, YIN Jun-xiang, RONG Jun

College of Life Science, Yangtze University, Jingzhou 434025,Hubei,China

Received:2022-01-19 Online:2022-05-25 Published:2022-06-14

摘要/Abstract

摘要： 制备禽腺病毒4型（FAdV-4）Fiber2蛋白的特异性单克隆抗体,将重组蛋白FAdV-4 Fiber2纯化后作为免疫原免疫BALB/c小鼠,分离脾脏中的淋巴细胞,与小鼠骨髓瘤细胞SP2/0进行融合,筛选得到10株能稳定分泌抗FAdV-4 Fiber2蛋白单克隆抗体的阳性杂交瘤细胞株1B5F1、1B5G11、1B5C1、1B5D2、1B5E3、1B5F3、1B5E4、1B5G4、1B5A8、1B5G8,取分泌抗体水平最高的1B5F1细胞株制备腹水,利用斑点杂交（Dot blot）和间接免疫荧光试验（IFA）进行特异性检测。结果表明,成功制备10株杂交瘤细胞株;制备的腹水能与FAdV-4 Fiber2蛋白及FAdV-4攻毒后的鸡肝组织产生特异性反应,证明成功制备了FAdV-4 Fiber2蛋白特异性单克隆抗体。

关键词: 禽腺病毒4型, Fiber2蛋白, 单克隆抗体

Abstract: The specific monoclonal antibody against avian adenovirus type 4 （FADV-4） Fiber2 protein was prepared. The FADV-4 Fiber2 protein was purified and immunized BALB/C mice as an immunogen. Lymphocytes were isolated from spleen and fused with mouse myeloma cells SP2/0. Ten hybridoma cell lines including 1B5F1, 1B5G11, 1B5C1, 1B5D2, 1B5E3, 1B5F3, 1B5E4, 1B5G4, 1B5A8 and 1B5G8 were screened for stable secretion of monoclonal antibody against FADV-4 Fiber2 protein. Ascites were prepared from 1B5F1 cell line with the highest antibody secretion level, and the specificity was detected by Dot blot and indirect immunofluorescence assay （IFA）. The results showed that 10 hybridoma cell lines were successfully prepared. The ascites could react specifically with FADV-4 Fiber2 protein and fADV-4 challenged chicken liver tissue, which proved that the specific monoclonal antibody of FADV-4 Fiber2 protein was successfully prepared.

Key words: serotype 4 fowl adenovirus, fiber2 protein, monoclonal antibody

中图分类号:

S831.7

谢明, 李国攀, 胡基雄, 张志翔, 艾笑扬, 陈帝斯, 李杨, 尹俊翔, 荣俊. 一株禽腺病毒4型Fiber2蛋白单克隆抗体的制备[J]. 湖北农业科学, 2022, 61(10): 121-126.

XIE Ming, LI Guo-pan, HU Ji-xiong, ZHANG Zhi-xiang, AI Xiao-yang, CHEN Di-si, LI Yang, YIN Jun-xiang, RONG Jun. Preparation of the monoclonal antibody against the serotype 4 fowl adenovirus fiber2 protein[J]. HUBEI AGRICULTURAL SCIENCES, 2022, 61(10): 121-126.

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一株禽腺病毒4型Fiber2蛋白单克隆抗体的制备

Preparation of the monoclonal antibody against the serotype 4 fowl adenovirus fiber2 protein

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