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目的 为提高抗猪囊虫单抗的特异性。方法 使用猪囊虫尿素溶性抗原 ( Ag- u)并用水溶性抗原 ( Ag- w)作对照分别免疫 BABL / c小鼠 ,取其脾细胞分别与 SP2 / 0骨髓瘤细胞融合 ,EL ISA法进行筛选。同时筛选出的阳性克隆扩大培养 ,取上清单抗分别与包虫、血吸虫、肝吸虫、肺吸虫、姜片虫和丝虫抗原进行交叉反应性测定。结果 Ag- u和 Ag- w的阳性克隆率分别为 13.5 % ( 13/ 96 )和 12 .5 % ( 12 / 96 ) ,特异性阳性克隆率分别为 30 .8% ( 4/ 13)和 8.3% ( 1/ 12 )。 SDS- PAGE分析 Ag- u和Ag- w的条带数分别为 9和 17,分子质量 >97.4ku的条带比率分别为 6 6 .7% ( 6 / 9)和 2 9.4% ( 5 / 17)。免疫组化显示来自 Ag-u的 2株单抗定位于囊虫的头节 ,而来自 Ag- w的 1株单抗定位于囊虫的囊壁。结论 Ag- u用于制备单抗的阳性克隆得率与Ag- w没有明显的差异 ,而特异性阳性克隆的得率是 Ag- w的 4倍 ,亦即 Ag- u不仅有良好的免疫原性 ,而且特异性更高
The purpose is to improve the specificity of anti-swine cysticercosis monoclonal antibody. Methods BABL / c mice were immunized with Agrobacterium tumefaciens U-Ag and immunized with water-soluble antigen (Ag-w). The spleen cells were fused with SP2 / 0 myeloma cells and ELISA method filter. The positive clones screened at the same time were expanded and cultured. The supernatant was used to determine the cross-reactivity with hydatid, schistosomiasis, liver fluke, paragonimiasis, ginger and silkworm antigens respectively. Results The positive clonality rates of Ag-u and Ag-w were 13.5% (13/96) and 12.5% (12/96) respectively, and the specific positive clonality rates were 30.8% (4/13) and 8.3 % (1/12). SDS-PAGE analysis showed that the bands of Ag-u and Ag-w were 9 and 17, respectively. The bands with molecular mass> 97.4ku were 66.7% (6/9) and 9.4% (5/17) ). Immunohistochemistry showed that two monoclonal antibodies from Ag-u were localized to the head section of the cysticercosis while one monoclonal antibody from Ag-w was localized to the cyst wall of the cysticercosis. Conclusion The yield of Ag-u for preparing monoclonal antibodies was not significantly different from that of Ag-w, while the yield of specific positive clones was 4-fold that of Ag-w, that is, Ag-u not only had good immunogen Sexuality, but also higher specificity