胃蛋白酶原Ⅱ单抗制备及夹心ELISA的初步建立  

Preparation of PepsinogenⅡMonoclonal Antibody and Development of Sandwich ELISA

作　　者：张红绪[1] 孙青芸[1] 王继创 李恒思 张怡青[1] 董彩文[3] 李玉林 刘旺根[3] 景建洲[3] 王云龙[4] 

机构地区：[1]河南师范大学,河南新乡453007 [2]河南生物工程技术研究中心,河南郑州450021 [3]郑州轻工业学院,河南郑州450021 [4]郑州职业技术学院,河南郑州450121

出　　处：《动物医学进展》2012年第4期56-59,共4页Progress In Veterinary Medicine

基　　金：河南省科技创新团队项目;郑州市科技创新团队项目

摘　　要：制备配对胃蛋白酶原Ⅱ(PGⅡ)单抗,建立人血清中胃蛋白酶原Ⅱ夹心ELISA检测方法。用胃蛋白酶原Ⅱ免疫Balb/c小鼠,制备免疫脾细胞,与SP2/0融合,用HAT培养基进行筛选培养,间接ELISA检测阳性克隆,对阳性孔进行多次单克隆化,选出效价高、分泌性能稳定的杂交瘤细胞,制备腹水并进行纯化。进行单抗配对,建立胃蛋白酶原Ⅱ双抗体夹心检测方法。获得1D8、1C6、2H11、2E3等4株杂交瘤,经配对试验,确定1D8、2H11可作为夹心ELISA检测PGⅡ的单抗。成功制备出配对单抗,初步建立了PGⅡ双抗体夹心ELISA方法。The purpose of this study was to prepare pepsinogen Ⅱ monoclonal antibody and establish the sandwich ELISA for serum PGⅡ.Balb / c mice were immunized by using pepsinogenⅡthe immune spleen cells were prepared and fused with SP2 / 0.After screening with HAT cultivation medium,indirect ELISA was used to screen positive clones.Stable hybridoma cells with high titer were obtained after cloning.Ascites were prepared and purified.Pepsinogen Ⅱ double antibody sandwich assay was established after antibody pairing test.Four hybridoma cell lines were obtained ： 1D8,1C6,2H11,2E3.1D8,2H11were determined as monoclonal antibody in the sandwich ELISA.The monoclonal antibodies were prepared,and the sandwich ELISA for detecting of PG Ⅱ was developed preliminary.

关 键 词：胃蛋白酶原 单克隆抗体 双抗体夹心ELISA 

分 类 号：S854.43[农业科学—临床兽医学]