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目的原核表达白细胞介素37(IL-37)及制备其多克隆抗体。方法 PCR扩增IL-37b成熟肽编码区基因,克隆至表达载体pET28a,转化大肠杆菌感受态细胞,IPTG诱导表达,Ni2+-NTA琼脂糖凝胶柱亲和层析纯化重组蛋白,以纯化的重组蛋白IL-37为免疫原免疫BALB/c小鼠制备其特异性抗体,用ELISA、Western blot法和免疫组织化学染色检测抗体的效价和特异性。结果原核表达了重组蛋白IL-37b成熟肽,并获得高效价的小鼠抗IL-37抗体,能特异性识别天然的IL-37抗原。结论成功制备效价高、特异性好的小鼠抗IL-37抗体。
Objective To prokaryotic express interleukin-37 (IL-37) and prepare its polyclonal antibody. Methods The coding region of mature peptide of IL-37b was amplified by PCR and cloned into expression vector pET28a. The recombinant plasmid was transformed into E.coli competent cells and induced by IPTG. Recombinant protein was purified by Ni2 + -NTA agarose gel column affinity chromatography. BALB / c mice were immunized with protein IL-37 to prepare their specific antibodies. The titer and specificity of the antibodies were tested by ELISA, Western blot and immunohistochemical staining. Results The prokaryotic recombinant protein IL-37b mature peptide was obtained and a high titer mouse anti-IL-37 antibody was obtained, which can specifically recognize natural IL-37 antigen. Conclusion The mouse anti-IL-37 antibody with high titer and specificity was successfully prepared.