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细粒棘球绦虫囊液McAb的制备及鉴定

PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODIES AGAINST ECHINOCOCCUS GRANULOSUS HYDATID CYST FLUID ANTIGEN

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【作者】 杨庆陵; 刘秉文; 缪德强; 王凤临;

【Author】 Yang Qingling;Liu Bingwen;Miao Deqiang;Wang Fenglin Departmt of Biochemistry

【机构】 华西医大生物化学教研室; 四川省寄生虫病防治研究所; 四川省计划生育干部管理学院;

【摘要】 作者用人源包虫囊液抗原免疫的BALB/c小鼠脾细胞与SP 2/0骨髓瘤细胞杂交,获得成功。融合率100%,抗体阳性率10.7%。经克隆化培养获得分泌抗人源细粒棘球绦虫抗原单克隆抗体的杂交瘤细胞5株,其染色体数为99~108。经鉴定其中3株(1D3、2D6、4B10)是特异针对细粒棘球绦虫抗原决定簇的,均属IgG1亚类;另外2株(4F4、1D1)则针对细粒棘球绦虫与其他几种寄生虫抗原的共同决定簇,它们分属IgM和IgG1类免疫球蛋白。

【Abstract】 McAbs were prepared by fusing SP2/0cells with spleen cells of BALB/c mice immu-nized with hydatid cyst fluid(HCF)antigen.Antibodies in hybrid culture snpernatantwere detected by ELISA, and positive hy-brids were cloned by technique of limitingdilutions. Numbers of chromosomes of fivehybridoma cell lines(1D1, 1D8, 2D6, 4B10,4F4) were demonstrated in the range of 99-108. The immunoglobulin class/subclass ofMcAb 4F4 was IgM and the others belong-ed to IgG1. IHA, ELISA and I-ELISA wereused for identifying titer and specificity offive McAbs. The titers of two McAbs (1D1,1D8) reached 1:106-107 in IHA and ELISA.Two of these McAbs (1D1, 4F4) with otherparasitic antigens revealed cross-reactions.This indicated that the McAbs 1D1 and 4F4were against common antigen determinersof different parasitic antigens and the McAbs1D8, 2D6, 4B10 against specific antigen de-terminers of HCF.

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