猪繁殖与呼吸综合征病毒核衣壳蛋白刺激小鼠产生抗体的研究

为比较传统免疫、脾内免疫、胶粒免疫和硝酸纤维素膜皮下包埋等4种方法制备的猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)核衣壳蛋白抗原免疫小鼠后刺激机体产生特异性抗体的水平,比较不同方法对于特定抗原刺激机体产生免疫应答的影响,筛选一种最有效的能够用于PRRSV核衣壳蛋白多克隆抗体和单克隆抗体制备的免疫方法。用PRRSV核衣壳蛋白作免疫原,分别用传统皮下免疫法、脾内免疫法、胶粒免疫和硝酸纤维素皮下包埋法免疫BABL/c小鼠,间接ELISA检测血清效价,用Western blotting检测其特异性。三免后,常规免疫小鼠血清效价为1.28×105,脾内免疫法为1.28×105,胶粒免疫为3.2×104,NC膜皮下包埋法为6.4×104。结果表明,4种免疫方法均获得了高滴度、高特异性的多克隆抗体,与传统免疫法相比,其他3种方法有其明显的优势,即节省抗原,其中脾内免疫法为优先选用的免疫方法,其免疫效果好,可节省大量抗原和时间。

The Study of the Antibody Produced by N Protein of Porcine Reproductive and Respiratory Syndrome Virus in Mice

The purpose of this study was to compare the levels of antibody titer to PRRSV and screen the best immunization method for PRRSV. The nucleocapsid protein of PRRSV was used as immunogen to immunize BABL/c mice by traditional immunization, NC filter immunization, intrasplenic immunization and gel particle containing special bands. Then the titer of positive sera was detected by indirect ELISA, and the specificity was analyzed by Western blotting. After the third immunization, the titers of serum in different mice groups immunized by the four groups were 1.28×105, 1.28×105, 3.2×104, and 6.4×104, respectively. The results showed that the four immunity methods had obtained high titers polyclonal antibody, comparing with traditional immunization, the other three methods had more advantage in conserving antigen. Intrasplenic immunization was the best immunization method, which had the best immunity effect，using the least antigen and the shortest time.