CD58抗体的制备及其检测方法的建立

Preparation of CD58 antibody and the establishment of its detection method

【目的】建立CD58特异性抗体的制备方法,及检测CD58抗体的间接ELISA方法。【方法】用CD58混合弗氏佐剂和兔淋巴细胞吸附CD58的方法免疫家兔获取CD58的抗体,双向免疫扩散试验检测抗体特异性,间接ELISA检测抗体水平。【结果】两种方法都能够使动物机体产生免疫应答;双向免疫扩散试验证实,弗氏佐剂法获得2种抗体,淋巴细胞吸附法制备抗原可获得1种抗体;间接ELISA检测法CD58的最适包被浓度为10.0μg/mL,最佳封闭液为10 g/L明胶,辣根酶标记的羊抗兔IgG工作浓度为1∶2 000。【结论】兔淋巴细胞吸附法可制备针对CD58的单一抗体,间接ELISA检测方法可以用于CD58抗体水平的检测。

[Objective] The study was to develop the method of preparation of specificity CD58 antibody with the purification CD58 from sheep,and to establish the indirect ELISA method of testing CD58 antibody. [Method] The unique antibody can be obtained by CD58 antigen and the mix Freund＇s adjuvant and CD58 antigen obtained by rabbit lymphocyte absorption. The double immunodiffusion tests the specificity, and indirect ELISA tests the antibody level. [Result] The immunologic response can be made by two methods. The double immunodiffusion test indicated that two antibodies can be obtained by the mix Freund＇s adjuvant, as one antibody can be obtained by another method. The most suitable concentration of coating antigen was 10.0/lg/mL,the most suitable block liquid 10 g/L Gelatin,and the working concentration of enzyme labeled goat-anti-rabbit antibody 1 ： 2 000. [Conclusion] The rabbit lymphocyte absorption can obtain the unique CD58 antibody,and the indirect ELISA can be used to detect the antibody level of CD58.