2型登革病毒NS1蛋白单克隆抗体的生物学特性研究

目的制备抗2型登革病毒NSl蛋白的单克隆抗体（mAb），并鉴定其特性。方法以重组NS1蛋白免疫Balb／c小鼠，采用杂交瘤技术制备抗M蛋白的mAb，采用间接ELISA方法和Westernblot进行mAb特异性鉴定；同时采用间接ELISA方法鉴定mAb相对亲和力。结果获得9株可分泌特异性mAb的杂交瘤细胞Ⅲ1A6andU13D2；相对亲和力均在10^6以上。Westernblot显示9株mAb能特异识别重组NS1蛋白。结论成功地制备出抗登革病毒2型病毒NS1蛋白的9株mAb，为建立快速特异检测登革病毒的实验方法提供了有力的工具。

Characterization of Monoclonal Antibody Against NSI Protein of Dengue Virus Serotype 2

Objective To prepare monoclonal antibodies(mAbs)against NS1 protein of dengue virus type 2(DV2),for the diagnosis of dengue virus infection.Methods After purification with Ni-NTA affinity column,the recombinant NS1 protein was used to immunize Balb/c mice.Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells.The affinity of the obtained mAbs was determined by indirect ELESA.The specificity of mAbs was tested by ELISA and Western blot analysis.Results Nine hybridomas were screened out,chromosome analysis revealed that hybridomas possessed the universal characteristics of the monoclonal hybridoma cells which secreted mAb.The relative affinity constants of mAbs were determined as over 106.Conclusions Specific mAbs against DV2 NS1 protein have been successfully prepared and identified.