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当前位置: 首页 > 文章 > 滑液支原体二氢硫辛酰胺乙酰转移酶单克隆抗体的制备及鉴定 中国兽医科学 2015 (8) 802-806
Position: Home > Articles > Preparation and identification of monoclonal antibodies against dihydrolipoamide acetyltransferase of Mycoplasma synoviae Chinese Veterinary Science 2015 (8) 802-806

滑液支原体二氢硫辛酰胺乙酰转移酶单克隆抗体的制备及鉴定

作  者：
郭小琴;包世俊;邢小勇;胡国明;薛慧文
单  位：
甘肃农业大学动物医学院陇西县动物疫病预防控制中心;甘肃农业大学动物医学院
关键词：
滑液支原体;单克隆抗体;杂交瘤;鉴定
摘  要：
为制备滑液支原体(MS)的单克隆抗体,将滑液支原体WVU 1853株全菌免疫的BALB/c小鼠脾细胞与小鼠骨髓瘤细胞SP2/0融合,在间接ELISA检测的基础上经有限稀释法克隆化,成功筛选出3株分泌抗MS WVU 1853株单克隆抗体的杂交瘤细胞融合株1B3、1E7及4E8。ELISA和Western-blot试验表明,这3株单克隆抗体可特异性地与滑液支原体WVU 1853株的32ku蛋白质结合,而与禽的其他几种常见病原微生物无任何反应。亚类鉴定结果表明,这3株单克隆抗体重链均为IgG1,轻链均为λ型。质谱分析结果表明,所获得的3株单克隆抗体均为抗滑液支原体二氢硫辛酰胺乙酰转移酶单克隆抗体。
译  名：
Preparation and identification of monoclonal antibodies against dihydrolipoamide acetyltransferase of Mycoplasma synoviae
作  者：
GUO Xiao-qin;BAO Shi-jun;XING Xiao-yong;HU Guo-ming;XUE Hui-wen;WEN Feng-qin;HU Yong-hao;XIANG Hai-tao;College of Veterinary Medicine,Gansu Agricultural University;Animal Disease Prevention and Control Center of Longxi County;
关键词：
Mycoplasma synoviae;;monoclonal antibody;;hybridoma;;identification
摘  要：
To prepare monoclonal antibodies(McAbs)against Mycoplasma synoviae,BALB/c mice were then immunized using whole cells of M.synoviae WVU 1853.The spleen cells from the immunized mice were fused with mouse myeloma SP2/0cells.There positive hybridoma cell lines secreting the McAbs against M.synoviae,namely 1B3,1E7 and 4E8,were selected by indirect enzyme-linked immunosorbent assay(ELISA)on the basis of limited dilution method.ELISA and Western-blot showed that the three McAbs were able to react with the protein with 32 ku in molecular weight specifically,but had no cross reactions with other avian pathogens.The immunoglobulin subtypes of the three McAbs were identified and the results showed that all the three McAbs were IgG1,and light chains were allλ.The mass spectrometry indicated that all the three McAbs were McAbs against the dihydrolipoamide acetyltransferase of M.synoviae.

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