两种抗原制备黄瓜花叶病毒多克隆抗体的比较研究

Comparative identification for two polyclonal antibodies to M Strain of Cucumber mosaic virus by using two kinds of antigens

以提纯的黄瓜花叶病毒M株系(CMV-M)病毒粒子以及体外原核表达的外壳蛋白(CMV-CP)为抗原免疫健康家兔,制备了CMV-M多克隆抗体。采用间接酶联免疫吸附试验检测其效价,并结合Western-blot比较了两种多克隆抗体的灵敏度。结果表明:制备的两种CMV-M多克隆抗血清效价均达到1:25600;检测病汁液的灵敏度方面,以病毒粒子为抗原的多克隆抗血清达到1:3200,明显优于以原核表达蛋白作为抗原的抗血清1:800,蛋白印迹着色前者在样品量倍比稀释后免疫条带清晰,后者条带模糊甚至消失。

Polyclonal antibodies were prepared by immunizing rabbits with purified CMV-M and the its coat protein expressed in E. coli respectively. The titers of two kinds of polyclonal antibody were tested using indirect enzyme- linked immunoadsordent assay （ID-ELISA）, and both antibodies＇ sensitivities were confirmed by western-blotting. The results showed that the titers of two kinds of anti-serum were the same 1： 25600, but the sensitivity detection results demenstrate that sensitivities of anti-serum prepared using CMV-M particles was 1：3200 and CMV-M -CP proteins as antigens was 1：800 when using plant rude extraction as test samples. In addition, the results of western-blot analysis showed that the three bands of anti-serum using CMV-M particles as antigens are more clearly than those of anti- serum using CMV-M -CP protein as antigens when the loading samples amount was double diluted into 3 different levels, which was consistent with the ID-ELISA results