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用水流动力学方法进行基因免疫,制备小鼠高效价多克隆抗体。采用水流动力学方法和肌肉注射pcDNA3.0/DR5免疫小鼠,分批在0、2、4、6周进行免疫,初次免疫8周后取鼠尾血用ELISA方法检测小鼠抗DR5抗体效价,用Westernblot、免疫组化染色法检测抗体的特异性。结果表明,ELISA结果显示末次免疫后水流动力学免疫组抗体滴度最高可达到为1∶102 400,肌肉注射免疫组抗体滴度最高可达1∶25 600。Western blot结果显示,小鼠抗DR5多克隆抗体识别Jurkat细胞的DR5蛋白条带。免疫组化结果表明,转染的DR5蛋白主要分布在食管癌细胞膜附近,与预期相符。用水流动力学的方法可以成功制备比肌肉注射更高效价的多克隆抗体。
Hydrodynamic method for gene immunization to prepare mouse high titer polyclonal antibody. The mice were immunized with hydrodynamic method and intramuscular injection of pcDNA3.0 / DR5. The mice were immunized at 0, 2, 4, and 6 weeks in batches. After 8 weeks of initial immunization, the mice were tested for anti-DR5 antibody by ELISA The specific antibody was detected by Western blot and immunohistochemistry. The results showed that the ELISA results showed that the highest titer of the antibody in the hydrodynamic group after the last immunization was 1:102 400, and the titer of the antibody in the intramuscular group was up to 1:25 600. Western blot results showed that mouse anti-DR5 polyclonal antibody recognized the DR5 protein band in Jurkat cells. The results of immunohistochemistry showed that the transfected DR5 protein mainly distributed near the cell membrane of esophageal cancer, which was in line with the expectation. Hydrodynamic methods allow the successful preparation of polyclonal antibodies that are more potent than intramuscular injections.