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目的应用HER-2 B细胞表位模拟小肽Mut与Hsc70的融合蛋白诱导抗小肽和抗HER-2的体液免疫反应。方法将Hsc70和Mut小肽序列定向插入pQE40载体中,构建pQE40-Hsc70-Mut原核表达质粒,并诱导表达、纯化融合蛋白Hsc70-Mut。常规免疫Balb/c小鼠,应用ELISA、Western blot检测小鼠血清中抗小肽Mut和抗HER-2的体液免疫反应。结果成功构建了pQE40-Hsc70-Mut原核表达质粒,Hsc70-Mut融合蛋白在大肠杆菌SG13009能够实现可溶性表达,应用Ni-NTA纯化可得到纯度>90%的融合蛋白;Hsc70-Mut免疫小鼠可产生抗Mut小肽的免疫反应,部分小鼠同时可产生抗HER-2的体液免疫反应。结论 Hsc70-Mut融合蛋白免疫小鼠可产生抗小肽和抗HER-2的体液免疫反应,为HER-2疫苗的研制奠定了一定基础,同时应用小肽融合蛋白免疫也为小肽免疫提供了一种新的免疫策略。
OBJECTIVE: To use HER-2 B cell epitope to mimic the fusion protein of small peptide Mut and Hsc70 to induce humoral immune response against anti-small peptide and anti-HER-2. Methods Hsp70 and Mut peptide sequences were inserted into pQE40 vector to construct the prokaryotic expression plasmid pQE40-Hsc70-Mut. The fusion protein Hsc70-Mut was purified and induced. Balb / c mice were routinely immunized, and the humoral immune responses of anti-small peptide Mut and anti-HER-2 in serum of mice were detected by ELISA and Western blot. Results The prokaryotic expression plasmid pQE40-Hsc70-Mut was constructed successfully. The Hsc70-Mut fusion protein was successfully expressed in Escherichia coli SG13009. The purity of fusion protein was over 90% by Ni-NTA purification. Hsc70-Mut immunized mice could produce Anti-Mut small peptide immune response, some mice can simultaneously produce anti-HER-2 humoral immune response. Conclusions Hsc70-Mut fusion protein immunized mice can produce humoral immune response against small peptides and anti-HER-2, which lays a foundation for the development of HER-2 vaccine. At the same time, immunization with small peptide fusion protein also provides small peptide immunization A new immunization strategy.