非洲猪瘟病毒pS273R蛋白水解酶单克隆抗体制备

doi:10.11843/j.issn.0366-6964.2021.03.028

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (3): 839-844.doi: 10.11843/j.issn.0366-6964.2021.03.028

非洲猪瘟病毒pS273R蛋白水解酶单克隆抗体制备

张萌^1,2, 刘英楠¹, 谢振华¹, 敖清莹¹, 吕璐¹, 于婉琪¹, 金文杰³, 秦爱建³, 扈荣良², 陈鸿军^1*

1. 中国农业科学院上海兽医研究所, 上海 200241;
2. 军事科学院军事医学研究院军事兽医研究所, 长春 130122;
3. 扬州大学兽医学院, 扬州 225009

收稿日期:2020-06-28 出版日期:2021-03-23 发布日期:2021-03-24
通讯作者: 陈鸿军,主要从事非洲猪瘟致病性研究,E-mail:vetchj@shvri.ac.cn
作者简介:张萌(1993-),女,河北廊坊人,硕士生,主要从事非洲猪瘟致病性研究,E-mail:1242715927@qq.com;刘英楠(1993-),男,山东威海人,博士生,主要从事非洲猪瘟致病性研究,E-mail:648152749@qq.com。
基金资助:
“十三五”国家重点研发专项资助项目（2017YFD0502300；2018YFC08400400）；自然科学基金联合基金项目重点支持项目-区域创新发展联合基金项目（U19A2039）

Preparation of Monoclonal Antibodies against African Swine Fever Virus pS273R Protease

ZHANG Meng^1,2, LIU Yingnan¹, XIE Zhenhua¹, AO Qingying¹, LÜ Lu¹, YU Wanqi¹, JIN Wenjie³, QIN Aijian³, HU Rongliang², CHEN Hongjun^1*

1. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China;
2. Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun 130122, China;
3. Yangzhou University, Yangzhou 225009, China

Received:2020-06-28 Online:2021-03-23 Published:2021-03-24

摘要/Abstract

摘要： 本研究旨在制备非洲猪瘟病毒（ASFV） S273R蛋白的特异性单克隆抗体。本研究以原核表达的非洲猪瘟病毒重组S273R蛋白免疫BALB/c小鼠，取其脾细胞与骨髓瘤细胞进行融合获得杂交瘤细胞。结果显示：基于纯化的S273R蛋白建立的间接ELISA方法对杂交瘤细胞进行筛选和亚克隆，获得了4株可稳定分泌抗ASFV S273R蛋白单克隆抗体的杂交瘤细胞株，并通过免疫印迹（Western blot）和免疫荧光试验（immunofluorescence assay，IFA）测定了其免疫特性。4株单克隆抗体亚类均为IgG1型，轻链均为κ链。本研究获得的非洲猪瘟病毒S373R蛋白单克隆抗体可为进一步研究ASFV pS273R的生物学功能及基因缺失毒株的鉴别诊断和疫苗开发提供技术储备。

关键词: 非洲猪瘟病毒, S273R蛋白, 原核表达, 单克隆抗体

Abstract: The objective of this study was to prepare monoclonal antibodies against the S273R protein of African swine fever virus (ASFV). BALB/c mice were immunized with purified recombinant S273R protein expressed by prokaryotic expression system and hybridoma cells were obtained by fusion of spleen cells and myeloma cells. An indirect ELISA based on the purified S273R protein was developed. Four hybridoma cell lines stably secreting MAbs against the S273R protein were harvested after screening and subcloning, and their sensitivity and specificity were detected by ELISA, Western blot, and immunofluorescence assay (IFA). Four MAbs heavy chains belong to IgG1, and light chains were κ. This research provided technical support for further study of biological functions of ASFV pS273R, as well as its gene-deletion based vaccine development.

Key words: African swine fever virus, S273R, prokaryotic expression, monoclonal antibody

中图分类号:

S852.659.1

张萌, 刘英楠, 谢振华, 敖清莹, 吕璐, 于婉琪, 金文杰, 秦爱建, 扈荣良, 陈鸿军. 非洲猪瘟病毒pS273R蛋白水解酶单克隆抗体制备[J]. 畜牧兽医学报, 2021, 52(3): 839-844.

ZHANG Meng, LIU Yingnan, XIE Zhenhua, AO Qingying, LÜ Lu, YU Wanqi, JIN Wenjie, QIN Aijian, HU Rongliang, CHEN Hongjun. Preparation of Monoclonal Antibodies against African Swine Fever Virus pS273R Protease[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 839-844.

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非洲猪瘟病毒pS273R蛋白水解酶单克隆抗体制备

Preparation of Monoclonal Antibodies against African Swine Fever Virus pS273R Protease

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