中国人兽共患病学报2017，Vol.33Issue(6)：481-485,512,6.DOI:10.3969/j.issn.1002-2694.2017.06.002
新城疫病毒核蛋白单克隆抗体的制备及双抗体夹心ELISA的初步建立
Development of monoclonal antibodies against nucleoprotein of Newcastle disease virus and establishment of a quantitative double-antibody sandwich ELISA for NDV antigen
摘要
Abstract
We developed the monoclonal antibodies against nucleoprotein (NP) of Newcastle disease virus (NDV),and established a double antibody sandwich ELISA method for quantitative determination of NP antigen of NDV (NDV NP ELISA).The recombination NP protein derived from strain F48E9 of NDV were prepared and used to immunize BLAB/c mice.The mouse splenic cells from immunized mice were fused with SP2/0 cells to generate monoclonal antibodies (mAb).The NDV NP specific mAbs were paired to establish a double antibody sandwich ELISA method.The performance of the NDV NP ELISA was evaluated,including specificity,sensitivity,precision,accuracy and linearity.The correlation between the ELISA and PFU virus titer was analyzed by regression analysis method.Two monoclonal antibodies 3C10 and 4E7 were selected to establish double antibody sandwich ELISA for NP antigen of NDV.The linearity and performance of the NDV NP ELISA was characterized.The detection linearity fell in the range of 0.015-0.250 μg/mL (R2 =0.997 4).The detection limit of the assay was 0.015 μg/mL.The recovery was between 88.4％ and 106.01％;the variation coefficient was below 3.4％.In testing of 50 NDV virus samples,this assay performed well and correlated comparably with PFU virus titer (R2 =0.920 9).The NDV NP ELISA for quantitative detection of NDV is a reliable quantifiable assay for detection of NDV NP protein;it provides a new approach for rapid and quantitative detection of Newcastle disease virus.关键词
新城疫病毒/核蛋白/单克隆抗体/定量检测/酶联免疫吸附测定Key words
Newcastle disease virus/NP protein/monoclonal antibody/quantitative assay/enzyme-linked immunosorbent assay (ELISA)分类
医药卫生引用本文复制引用
王明睿,钟建平,李睿,王国松,陈毅歆..新城疫病毒核蛋白单克隆抗体的制备及双抗体夹心ELISA的初步建立[J].中国人兽共患病学报,2017,33(6):481-485,512,6.基金项目
国家自然科学基金(31670934):基于广谱中和单抗的通用型流感疫苗设计及其结构基础研究 Supported by the National Natural Science Foundation of China (Grant No.31670934):The design of universal influenza vaccine based on broadly neutralizing monoclonal antibody and its structural study. （31670934）
