抗葡萄病毒A单克隆抗体的制备、特性鉴定及应用

Production, characterization and use of monoclonal antibodies to grapevine virus A.

作者信息

Boscia D, Aslouj E, Elicio V, Savino V, Castellano M A, Martelli G P

机构信息

Dipartimento di Protezione delle Piante, Università degli Studi, Bari, Italy.

出版信息

Arch Virol. 1992;127(1-4):185-94. doi: 10.1007/BF01309583.

DOI:10.1007/BF01309583

PMID:1280942

Abstract

Four stable hybridoma cell lines secreting monoclonal antibodies to grapevine closterovirus A (GVA) were obtained by fusing spleen cells of immunized BALB/c mice with mouse myeloma cell line Sp2/0-Ag 14. In ELISA all MAbs reacted with virus in leaf extracts from Nicotiana benthamiana, glass-house-, field-, or in vitro-grown grapevines, or with cortical shavings from mature grape canes. In IEM tests, only one of the MAbs (PA3.F5) decorated virus particles on the entire surface. This MAb was likely induced by a surface antigenic determinant, whereas the other three MAbs (PA3.D 11, PA3.C 6, and PA3.B 9) were originated by cryptotopes. Coupling polyclonal antibodies for coating protein A-sensitized plates, and monoclonal antibody conjugates for antigen detection, gave highly efficient and reproducible results for identification of GVA in field-grown grapevines.

摘要

通过将免疫的BALB/c小鼠的脾细胞与小鼠骨髓瘤细胞系Sp2/0-Ag 14融合，获得了四种分泌抗葡萄藤褪绿斑驳病毒A（GVA）单克隆抗体的稳定杂交瘤细胞系。在酶联免疫吸附测定（ELISA）中，所有单克隆抗体都能与来自本氏烟草、温室、田间或离体培养葡萄藤的叶片提取物中的病毒发生反应，或与成熟葡萄茎的皮层刨花发生反应。在免疫电镜（IEM）测试中，只有一种单克隆抗体（PA3.F5）能在病毒粒子的整个表面进行标记。这种单克隆抗体可能是由表面抗原决定簇诱导产生的，而其他三种单克隆抗体（PA3.D 11、PA3.C 6和PA3.B 9）则是由隐蔽表位产生的。将用于包被蛋白A致敏板的多克隆抗体与用于抗原检测的单克隆抗体结合物相结合，在鉴定田间种植葡萄藤中的GVA时，能产生高效且可重复的结果。