[百合无症状病毒单克隆抗体的制备及其在百合病毒检测中的应用]

[Production of monoclonal antibodies to Lily symptomless virus and application in lily virus detection].

作者信息

Wu Jian-Xiang, Liu Cheng-Ke, Hong Jian, Liu Wen-Hong, Ye Mei-Qing

机构信息

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.

出版信息

Wei Sheng Wu Xue Bao. 2005 Aug;45(4):580-3.

PMID:16245875

Abstract

Four hybridoma cell lines, 2A2, 5H9, 5H2 and 5E12, secreting monoclonal antibodies (MAbs) against Lily symptomless virus (LSV) were produced by fusing mouse myeloma cells (SP2/0) with spleen cells from BALB/C immunized by the LSV particles. The four MAbs could specifically react with LSV. The titres of ascitic fluids of the four MAbs are up to 10(-6) in ELISA. Isotypes and subclasses of 5H9 and 5E12 belong to IgG1 while those of 2A2 and 5H2 belong to IgG3. Isotypes of light strains of the four MAbs all belong to kappa. The four MAbs were used in antigen-coated plate (ACP)-ELISA for LSV detection, and ACP-ELISA could successfully detect 1.8 ng of purified LSV or virus in plant sap diluted 1:300. The presence of LSV in field lily tissues was investigated with ACP-ELISA.

摘要

通过将小鼠骨髓瘤细胞（SP2/0）与经百合无症状病毒（LSV）颗粒免疫的BALB/C小鼠的脾细胞进行融合，产生了4株分泌抗LSV单克隆抗体（MAb）的杂交瘤细胞系，分别为2A2、5H9、5H2和5E12。这4种单克隆抗体能与LSV特异性反应。在ELISA中，这4种单克隆抗体腹水的效价高达10⁻⁶。5H9和5E12的重链亚型属于IgG1，而2A2和5H2的重链亚型属于IgG3。这4种单克隆抗体轻链的亚型均属于kappa。将这4种单克隆抗体用于抗原包被板（ACP）-ELISA检测LSV，ACP-ELISA能够成功检测到1.8 ng的纯化LSV或稀释1:300的植物汁液中的病毒。利用ACP-ELISA对田间百合组织中的LSV存在情况进行了调查。