禽白血病病毒p27蛋白单克隆抗体制备及抗原表位的鉴定  

Preparation of Monoclonal Antibodies against p27 Protein of Avian Leukemia Virus and Identification of Antigenic Epitopes

作　　者：陈桂娥 容芳 苟鑫 孙荣航 李作生 陈瑞爱 CHEN Gui-e;RONG Fang;GOU Xin;SUN Rong-hang;LI Zuo-sheng;CHEN Rui-ai(Zhaoqing Branch Center of Guangdong Laboratory for Lingnan Modern Agricultural Science and Technology,Zhaoqing,Guangdong,526238,China;College of Life Sciences,South China Agricultural University,Guangzhou,Guangdong,510642,China;College of Veterinary Medicne,South China Agricultural University,Guangzhou,Guangdong,510642,China;Zhaoqing Institute of Biotechnology Co.,Ltd.,Zhaoqing,Guangdong,526238,China;Zhaoqing Dahuanong Biological Medicine Co.,Ltd.,Zhaoqing,Guangdong,526238,China)

机构地区：[1]岭南现代农业科学与技术广东省实验室肇庆分中心,广东肇庆526238 [2]华南农业大学生命科学学院,广东广州510642 [3]华南农业大学兽医学院,广东广州510642 [4]华农(肇庆)生物产业技术研究院有限公司,广东肇庆526238 [5]肇庆大华农生物药品有限公司,广东肇庆526238

出　　处：《动物医学进展》2025年第1期23-28,共6页Progress In Veterinary Medicine

基　　金：岭南现代农业科学与技术广东省实验室肇庆分中心自主项目(P20211154-0201);肇庆市重点领域研发计划“揭榜制”项目(2021C001)。

摘　　要：为制备禽白血病病毒(ALV)p27蛋白特异性单克隆抗体,利用原核表达系统表达并纯化ALV p27蛋白,将其作为免疫原免疫注射Balb/c小鼠,用杂交瘤细胞融合和ELISA等筛选出阳性单克隆抗体细胞,对制备的单克隆抗体的抗体亚型、效价及特异性进行鉴定,将目的蛋白截短成4段,初步鉴定单克隆抗体识别的抗原表位区域。结果表明,共筛选出4株单克隆细胞,其抗体亚类皆为IgG1型,其中3A1抗体效价为1∶64000,3B2为1∶256000,5F1为1∶128000,5G2为1∶8000;4株单克隆抗体识别的抗原表位在1-60 aa之间。研究结果为进一步建立ALV相关免疫学检测方法提供了重要的材料。To prepare avian leukemia virus(ALV)p27 protein specific monoclonal antibody,the experiment was used prokaryotic expression system to express and purify ALV p27 protein,using it as an immunogen to immunize Balb/c mice.Through hybridoma cell fusion and ELISA,the positive monoclonal antibody cells was screened.The subclass,titer and specificity of monoclonal antibodies was detected.The target protein was truncated into four segments for preliminary identification of antigen epitope recognized by monoclonal antibody.The results showed that four monoclonal cell lines were selected and their antibody subclass were IgG1.The antibody titers were 1∶64000 for 3A1 cell line,1∶256000 for 3B2 cell line,1∶128000 for 5F1 cell line,and 1∶8000 for 5G2 cell line.The epitope sequences recognized by four cell line were between 1-60 amino acids.The results provided an important material for the further development of immunological detection methods for ALV.

关 键 词：禽白血病病毒 P27蛋白 单克隆抗体 抗原表位 

分 类 号：S852.659.3[农业科学—基础兽医学] S858.31[农业科学—兽医学]