旋毛虫53kuES重组蛋白单克隆抗体的制备及其免疫金标试纸条的研制

为建立一种简易、快速检测动物血清中的旋毛虫排泄分泌(Excretory-secretory,ES)抗原的胶体金免疫层析试纸法,本研究选用旋毛虫53 ku ES重组蛋白免疫BALB/c小鼠,应用杂交瘤细胞技术制备单克隆抗体(MAb).利用辛酸-饱和硫酸铵法和亲和层析柱联合法纯化MAb及兔抗旋毛虫53 ku ES多克隆抗体,利用枸橼酸三钠还原法制备的胶体金颗粒标记MAb,制备免疫金标试纸条.结果表明,获得的2株IgG1型杂交瘤细胞系4D10和5A2能够稳定分泌抗体,且效价高；胶体金标记MAb 4D10所制备的试纸条具有重复性好、灵敏性和特异性高的优点；该试纸条4℃条件下可密封保存6个月以上.

Establishment of gold immunochromatography assay for detecting Trichinella spiralis with a monoclonal antibody against 53 ku ES recombination protein

To develop a rapid,simple gold immunochromatographic assay for the serological detection of the level of the excretory-secretory(ES) antigen of Trichinella,the monoclonal antibody(MAb) against 53 ku ES protein of Trichinella spiralis was prepared through the hybridoma technology,and the gold particle was coated with the MAb prepared by mixing Tri-sodium citrate with aqueous gold chloride to prepared the detection strip.In the strip,the MAb labelled colloidal gold was used as the detector,and the polyclonal anti-53 ku ES antibodies and goat anti-mouse IgG antibody were blotted on the nitrocellulose membrane for the test and control lines,respectively.The experimental results showed two hybridoma cell lines named 4D10 and 5A2 were obtained,which could stably secrete high level MAb.The immunochromatographic strip was shown to be of high specificity,sensitivity and reproducibility.At the temperature of 4 degree,the strip could also detect samples when sealed for six months.