植物抗体基因工程：I．分泌马铃薯Y病毒中和抗体杂交瘤细胞系的筛选

Antibody Genetic Engineering in Plant :I.Selection of a Hybridoma Cell Line Secreting Monoclonal Antibody Against Potato Virus Y

酶联免疫吸附试验证实，Ｄ７３、Ｄ９０和Ａ６９三个杂交瘤细胞系产生的单克隆抗体均能够与马铃薯Ｙ病毒株系ＰＶＹ￣ｏ、ＰＶＹ￣Ｎ和ＰＶＹ￣ｃ发生反应，但以Ｄ７３的亲和性为最强。进而进行的侵染抑制试验也证实经Ｄ７３单克隆抗体处理后的病毒接种物，其在‘Ａ６’（ＳｏｌａｎｕｍｄｅｍｉｓｓｕｍｘＳ．ｔｕｂｅｒｏｓｕｍ）离休叶片引起枯斑的能力显著地受到抑制。因此，选用Ｄ７３杂交瘤细胞系作为分离马铃薯Ｙ病毒中和抗体基因的材料，对其分泌的抗体轻链和重链的亚类进行鉴定，结果发现组成该单克隆抗体分子的轻链和重链分别为Ｋ链（ｋａｐｐａ）和γ＿１链。

By an ELISA assay,D73,a mouse hybridoma cell line,was selected for the production of monoclonalantibcaly which reacts with the three strains of potato virus Y──PVY°,PVYN and PVYc,which has thehighest affinitiy compared with other two hybridoma cell lines. Detached leaves of‘A6’(Solanum demissum x S. tuberosum ) were inoculated,the left half with a PVYo , PVYN or PVYc strain,and the right half with thesame inoculum incubated with the diluted different concentrations of the purified monoclonal antibody(original 1mg/ml)from D73 overnight prior to inoculation. We found that the number of necrotic locallesions was obviously reduced on the halves of the detached leaves treated by the mon oclonal antibody.Identification of heavy and light chain class and subclass of the monoclonal antibodies was carried out bydirect enzyme-linked immunosorbent assay with goat anti-mouse kappa,lameda, 1,γ2, γ3 and μ-chain-specific antibodies conjugated to alkaline phosphatase separately. The result showed that the antibodyconsists of kappa,light chains and γ1 heavy chains.