靶向DC-SIGN的Le(x)-OVA抗原的制备

Preparation of Le(x)-OVA antigen targeting to DC-SIGN

目的制备用于靶向DC-SIGN受体的Le(x)寡糖介导的靶向抗原。方法利用Su lfo-SMCC异源双功能交联试剂交联链霉亲和素(streptavid in,SA)和模式抗原卵白蛋白(ovalbum in,OVA),再利用SA与生物素高亲和力结合的特性,将SA-OVA复合物与生物素修饰的Le(x)寡糖反应,由此获得由Le(x)寡糖介导的靶向抗原Le(x)-OVA复合物。结果应用SDS-PAGE蛋白质电泳和W estern免疫印迹方法证实SA与OVA蛋白成功交联,ELISA方法证实SA-OVA复合物中的SA仍具有与生物素反应的功能,根据SA-OVA复合物与生物素修饰的Le(x)寡糖饱和反应的比例,获得所需要的靶向抗原Le(x)-OVA复合物。结论获得Le(x)寡糖介导的靶向抗原Le(x)-OVA复合物,为进一步研究经由DC-SIGN靶向后的抗原特异性免疫应答奠定了基础。

Objective To make Le（x）-OVA antigen targeting to DC-SIGN mediated by Le（x） oligosaccharide. Methods Utilizing Sulfo-SMCC as cross-linker, the streptavidin was conjugated to OVA, and the SA-OVA were linked with Le（x） sugars through interaction of SA and biotin, thus Le（x）-OVA antigen was obtained. Results SA was successfully conjugated to OVA by SDS-PAGE and Western blotting, and SA still had the capacity to react with biotin after conjugated to OVA, as shown in ELISA assay. According to ratio of saturated reaction between SA conjugated to OVA and biotin labeled to Le（x） sugars, the Le（x）-OVA antigen were obtained. Conclusion The antigen targeting to DC-SIGN, Le（x）-OVA, was obtained, which prepared the materials for further research of immune response induced by Le（x）-OVA.