[1]屈贵蜀,何 晴,严梦涵,等.番鸭呼肠孤病毒p10.8蛋白单克隆抗体的制备[J].福建农林大学学报(自然科学版),2021,50(05):655-660.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2021.05.012]
 QU Guishu,HE Qing,YAN Menghan,et al.Preparation of monoclonal antibody against muscovy duck reovirus p10.8 protein[J].,2021,50(05):655-660.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2021.05.012]
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番鸭呼肠孤病毒p10.8蛋白单克隆抗体的制备()
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福建农林大学学报(自然科学版)[ISSN:1671-5470/CN:35-1255/S]

卷:
50卷
期数:
2021年05期
页码:
655-660
栏目:
出版日期:
2021-09-25

文章信息/Info

Title:
Preparation of monoclonal antibody against muscovy duck reovirus p10.8 protein
文章编号:
1671-5470(2021)05-0655-06
作者:
屈贵蜀 何 晴 严梦涵 陆芍华 黄瑞玲 彭瑶顺 庄许诺 许丽惠 王全溪
[福建农林大学动物科学学院(蜂学学院),福建 福州 350002]
Author(s):
QU Guishu HE Qing YAN Menghan LU Shaohua HUANG Ruiling PENG Yaoshun ZHUANG Xunuo XU Lihui WANG Quanxi
[College of Animal Sciences(College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China]
关键词:
番鸭呼肠孤病毒 p10.8蛋白 单克隆抗体
Keywords:
muscovy duck reovirus p10.8 protein monoclonal antibody
分类号:
S852.4+3
DOI:
10.13323/j.cnki.j.fafu(nat.sci.).2021.05.012
文献标志码:
A
摘要:
为制备番鸭呼肠孤病毒(MDRV)p10.8蛋白的单克隆抗体(mAb),利用原核表达系统表达p10.8蛋白,采用Ni-NTA亲和层析法对其进行纯化,选择BALB/c小鼠进行免疫.4免后取小鼠脾细胞与骨髓瘤细胞SP2/0进行细胞融合,并以纯化的重组p10.8蛋白作为抗原包被,用间接ELISA法筛选到1株针对MDRV p10.8蛋白的能稳定分泌mAb的杂交瘤细胞株,命名为H3-7B株.用Western blotting法进行检测,以H3-7B株上清为一抗,分别与原核、真核表达系统获得的重组p10.8蛋白反应,结果均呈阳性.用抗体亚类试剂盒进行抗体亚类测定,显示H3-7B株mAb亚型为IgG2a类.用MDRV、禽腺病毒、鸭坦步苏病毒3种病毒与H3-7B株mAb进行Western blotting鉴定反应,特异性反应结果表明,H3-7B株mAb与MDRV反应呈阳性,特异性较好.结论:本试验成功制备了抗MDRV p10.8蛋白的mAb.开放科学(资源服务)
Abstract:
In order to prepare monoclonal antibody(mAb)of muscovy duck reovirus(MDRV)p10.8 protein, p10.8 protein was expressed by prokaryotic expression and purified using Ni-NTA affinity chromatography. Then it was subcutaneously injected into BALB/c mice. After the 4th subcutaneous injection, the spleen of the mice was removed and fused with myeloma cell SP2/0. The purified recombinant protein was used as the antibody coating, and the hybridoma cell strain stably producing mAb against MDRV p10.8 protein was screened by indirect enzyme-linked immunosorbent assay and was named as H3-7B. The recombinant proteins p10.8, produced from prokaryotic expression and eukaryotic expression, were both tested positive by Western blotting with the supernatant of H3-7B strain being the primary antibody. The antibody subclass was identified as IgG2a using antibody subclass kit. Western blotting analysis of MDRV, avian adenovirus and duck tembusu virus and H3-7B strain mAb showed positive reaction of MDRV and good specificity. Therefore, mAb against MDRV p10.8 was successfully produced in this study.

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相似文献/References:

[1]蔡一龙,庄育彬,周五朵,等.MDRV P10基因的克隆、分析及 蛋白质结构预测[J].福建农林大学学报(自然科学版),2015,44(06):606.[doi:10.13323/j.cnki.j.fafunat.sci.2015.06.008]
 CAI Yi-long,ZHUANG Yu-bin,ZHOU Wu-duo,et al.Cloning, sequence analysis and predicted protein construction of MDRV P10 gene[J].,2015,44(05):606.[doi:10.13323/j.cnki.j.fafunat.sci.2015.06.008]

备注/Memo

备注/Memo:
收稿日期:2020-11-16 修回日期:2021-01-08 基金项目:福建省自然科学基金资助项目(2013J01069、2019J0658). 作者简介:屈贵蜀(1995-),女.研究方向:预防兽医.Email:18104476289@163.com.通信作者王全溪(1978-),男,副教授,硕士生导师,博士.研究方向:宿主与病原互作.Email:wqx608@126.com.
更新日期/Last Update: 1900-01-01