人参皂苷Rb1单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibody against ginsenoside Rb1

作　　者：赵寿经[1] 侯春喜[1] 钱延春[1] 徐立新[1] 孙莉丽[1] 

机构地区：[1]吉林大学生物与农业工程学院,长春130022

出　　处：《吉林大学学报（工学版）》2007年第1期245-248,共4页Journal of Jilin University:Engineering and Technology Edition

基　　金：吉林省科技发展计划项目(20060546)

摘　　要：用人参皂苷Rb1分别与牛血清白蛋白（BSA）和卵清蛋白（OVA）反应合成了交联抗原GRb1-BSA（GRb1与BSA的结合比为10：1）和GRb1—OVA（GRb1与OVA的结合比为8：1）。以GRb1—BSA为免疫原，分5次免疫3只BAuyC小鼠，取免疫脾细胞与小鼠骨髓瘤细胞进行融合，Hat筛选，有限稀释法克隆。建立分泌单克隆抗体的杂交瘤细胞系。采用ELISA（酶联免疫吸附测定）间接法和竞争法检测抗体的特异性。结果获得了一只小鼠分泌抗体效价达1：16000，并获得两株分泌GRb1的单克隆抗体的细胞系，分别命名为1F7和1G3，单抗检测显示在0．01～1μg／mL呈线性关系，GRb1的检测范围达50～350ng／mL，两株单抗针对相同的抗原决定簇，制备的单克隆抗体可用于GRb1含量的检测和分离。Cross-linking antigens GRb1-BSA （ratio 10 ： 1） and GRb1-OVA （ratio 8 ： 1） were produced by the reactions of ginsenoside Rb1 with serum albumin （BSA） and ovalbumin （OVA） respectively. Three BALB/C mice were immunized five times with GRb1-BSA as the immunogen. Using polyethylene glycol （PEG） method, the immunized splenocytes were isolated and fused with a hypoxanthine-aminopterin-thymidine-sensitive mouse myeloma cell line, SP2/0. Hybridomas that produce monoclonal antibody （McAb） reactive to GRb1 were cloned by limiting dilution method. Hybridoma cell line that secretes McAb was obtained. Indirect enzyme-linked immunosorbent assay （ELISA） was applied to determine the titer of antisera and competitive ELISA was used to detect McAb. As results, the titer of the antisera is over 1 ： 16 000 by ELISA, two cell lines （1F7, 1G3） that secrete McAb are obtained, McAb detection shows linear relation in the range from 0.01 g/mL to 1 μg/mL. The full range of GRb1 detection is from 50 ng/mL to 350 ng/mL by competitive ELISA. The two McAbs are contraposed the same determining cluster and can be used to establish ELISA for GRb1 content detection and GRb1 isolation.

关 键 词：生物化学工程 单克隆抗体 人参皂苷RB1 酶联免疫吸附测定(ELISA) 

分 类 号：S567.51[农业科学—中草药栽培]