Zhang Jun, Zhang Juan, Tang Ni, Zhang Bing-qiang, Chen Wei-xian, Huang Ai-long, Zheng Jian
Key Laboratory of Infectious Diseases of the Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 May;22(3):374-6.
To prepare monoclonal antibody (mAb) against Vibrio cholerae O139, which would be used as the gold colloidal reagent strip for rapid detection of O139, and to determine its biological characterization.
BALB/c mice were immunized by inactivated Vibrio cholerae O139. Anti-O139 mAbs were prepared by using hybridoma technique. The specificity of mAb was determined by indirect ELISA and Western blot. The indirect ELISA was used to identify Ig subgroup, detect its titer in ascites and relative affinity, and analyze antigen-binding epitope of mAbs.
Two hybridoma cells (O4D7 and O4D10), secreting anti-O139 mAbs were obtained. The Ig subgroups of O4D7 and O4D10 were IgG2b and IgG3, respectively. The mAb's titer in ascites was 1:10(7). The relative affinity of mAb O4D7 was more than 10(5) and that of O4D10 was more than 10(4). The result of additive ELISA showed that two mAbs could recognize different antigen epitopes.
The successful preparation of two anti-O139 mAbs provides a powerful tool for the development of a rapid method of detecting Vibrio cholerae O139 infection.
制备抗霍乱弧菌O139单克隆抗体，用于霍乱弧菌O139快速检测的胶体金试剂条，并确定其生物学特性。
用灭活的霍乱弧菌O139免疫BALB/c小鼠，采用杂交瘤技术制备抗O139单克隆抗体。通过间接ELISA和Western blot法测定单克隆抗体的特异性。采用间接ELISA法鉴定Ig亚类，检测其腹水效价和相对亲和力，并分析单克隆抗体的抗原结合表位。
获得两株分泌抗O139单克隆抗体的杂交瘤细胞（O4D7和O4D10）。O4D7和O4D10的Ig亚类分别为IgG2b和IgG3。腹水效价为1:10(7)。单克隆抗体O4D7的相对亲和力大于10(5)，O4D10的相对亲和力大于10(4)。竞争ELISA结果显示，两株单克隆抗体可识别不同的抗原表位。
成功制备两株抗O139单克隆抗体，为建立霍乱弧菌O139感染快速检测方法提供了有力工具。
Zotero 插件