抗α-苦瓜素单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibody Against α-momorcharins

作　　者：孟尧[1,2] 姚兴川[3] 王殊睿[3] 刘敏[3] 王红宁[3] 孟延发[3] 赵晓军[1,2] 

机构地区：[1]四川大学纳米生物医学技术与膜生物学研究所 [2]四川大学生命科学学院,成都610064 [3]四川大学生命科学学院/四川省动物疾病预防和食品安全重点实验室,成都610064

出　　处：《生物医学工程学杂志》2011年第6期1181-1184,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30770232)

摘　　要：用α-苦瓜素(α-MMC)免疫小鼠(BALB/c),分离免疫脾细胞,采用聚乙二醇(PEG)法将免疫脾细胞与小鼠骨髓瘤细胞(SP2/0)融合,筛选出分泌抗α-MMC素的单克隆抗体(McAb)杂交瘤细胞株;采用小鼠体内诱生法制备腹水,Mabselect亲和层析纯化McAb,所得McAb经亚类鉴定为IgG2b亚类,通过间接酶联免疫吸附试验(ELISA)和Western blot检测,所得McAb与α-MMC特异反应,与相同来源的MAP30无交叉反应,为深入研究α-MMC的结构与功能提供了检测手段。Three BALB/c mice were immunized four times with α-momorcharins （α-MMC）. Using polyethy-lene glycol（PEG） method, the immunized splenoeytes were fused with SP2/0 cells. One strain of hybridoma cells was obtained which secrete antibodies against α-MMC. To get ascites, the hybridoma cells were injected into the abdominal cavity of mice. The antibodies were purified from aseites. Indirect enzyme linked immunosorbent assay （ELISA） and Western blot assay were applied to determine the specifity of the monoclonal antibody （MeAb）. The xesults showed that the McAb was specific to α-MMC without detectable cross-activity with MAP30. The MeAb provided detecting method for further research of the structure and function of α-MMC.

关 键 词：α-苦瓜素 酶联免疫吸附试验 单克隆抗体 亲和层析 WESTERN BLOT检测 

分 类 号：R392[医药卫生—免疫学]