水牛黏液病毒抗性蛋白_1的单克隆抗体

Q2 Veterinary Journal of Buffalo Science Pub Date : 2020-02-20 DOI:10.6000/1927-520x.2020.09.03
Dam Van Phai, V. Sophie, Dubois Axel, Bui Tran Anh Dao, Desmecht Daniel
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引用次数: 0

摘要

:利用大肠杆菌表达系统成功表达了重组bubaline (Bubalus bubalis)黏液病毒抗性蛋白1 (bbMx1)。免疫和细胞融合后,建立了一组10只产生bbMx1单克隆抗体的小鼠杂交瘤。采用间接ELISA、western blot和免疫细胞荧光染色对10个对应的单抗进行进一步鉴定。8个单克隆抗体分别为11C7、RD5、NF3、9D1、FD4、PE6、11A7和10F5,在三种格式下均显示出结合能力和特异性。此外,将NF3(用于捕获)与RD5(用于检测)、9D1与RD5、FD4与11C7或PE6与11C7结合,在原型三明治- elisa中产生强信号。结果表明,本文开发和表征的单克隆抗体为开发用于检测水牛病毒感染的诊断工具提供了一个很好的起点,无论是使用免疫印迹法、免疫细胞染色法还是三明治elisa法。
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Monoclonal Antibodies Specific to Water Buffalo (Bubalus bubalis) Myxovirus Resistance Protein_1
: Recombinant bubaline ( Bubalus bubalis ) myxovirus resistance protein 1 (bbMx1) was successfully expressed by an Escherichia coli expression system. After immunization and cell fusion, a set of ten mouse hybridomas producing mAbs to bbMx1 was established. The ten corresponding mAbs were further characterized using indirect ELISA, western blot analysis and immunocytofluorescent staining. Eight mAbs, designated 11C7, RD5, NF3, 9D1, FD4, PE6, 11A7 and 10F5, displayed binding abilities and specificity in the three formats. Moreover, combining NF3 (for capture) with RD5 (for detection), 9D1 with RD5, FD4 with 11C7 or PE6 with 11C7 generated a strong signal in a prototype sandwich-ELISA. The results suggest that the mAbs developed and characterized here provide an excellent starting point for developing diagnostic tools aimed at detecting viral infections in the water buffalo, whatever using immunoblotting, immunocytostaining or sandwich-ELISA.
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来源期刊
Journal of Buffalo Science
Journal of Buffalo Science Veterinary-Veterinary (all)
CiteScore
2.40
自引率
0.00%
发文量
9
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