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目的:以His-CecropinXJ融合蛋白、pcDNA3.0-cecropinXJ重组质粒为免疫原,分别采用蛋白皮下免疫、DNA肌肉免疫及使用活体基因导入仪肌肉免疫BALB/c小鼠,制备新疆家蚕抗菌肽的多克隆抗血清,并评价不同免疫方案对抗原免疫原性的影响。方法:采用间接ELISA测定免疫血清中抗新疆家蚕抗菌肽IgG抗体的滴度及亚型分类。用Western blot检测各免疫方案制备的抗血清,与新疆家蚕抗菌肽CecropinXJ抗原结合的特异性。结果:不同免疫方案均可诱导产生针对新疆家蚕抗菌肽不同滴度的多克隆抗血清。使用活体基因导入仪免疫后,可诱导高滴度的抗体产生。结论:以His-CecropinXJ蛋白免疫BALB/c小鼠后,可诱导高滴度的IgG1抗体产生;以pcDNA3.0-cecropinXJ质粒免疫后,可诱导针对IgG2a的抗体产生。
OBJECTIVE: To prepare the antimicrobial peptides of Bombyx mori in Xinjiang by using His-CecropinXJ fusion protein and pcDNA3.0-cecropinXJ recombinant plasmid as the immunogen, and immunizing the BALB / c mice respectively with protein subcutaneous immunization, DNA muscle immunization and muscle biopsy instrument Antisera were cloned and the effects of different immunization protocols on the immunogenicity of the antigen were evaluated. Methods: Indirect ELISA was used to determine the titer and subtype of anti-human IgG anti-Xinjiang antibody in serum. Western blot was used to detect the specificity of the antisera prepared by each immunization protocol and the antimicrobial peptide CecropinXJ antigen of Xinjiang silkworm. Results: Different immunization regimens could induce polyclonal antiserum against different titer of antimicrobial peptide in Xinjiang silkworm. After immunization with a live gene introducer, high titers of antibody production can be induced. Conclusion: The immunization of BALB / c mice with His-CecropinXJ protein can induce the production of high-titer IgG1 antibody. After immunization with pcDNA3.0-cecropinXJ plasmid, the antibody against IgG2a can be induced.