Preparation of HER-2 vaccine and immune response in mice
AIM: To prepare the vaccine of HER-2 antigen. METHODS: cDNA of HER2 extracellular domain was prepared through RT-PCR from HER-2 + SKBR cell line, cloned into the multiple cloning site of pQE-30 and expressed in E. coli DH5α. The expressed protein was purified through Ni-NTA resin affinity chromatography. Eluted protein was refolded by different concentrations of urea and then detected by Western Blotting. The protein immunization antibody detection in immunized mice was examined by ELISA. RESULTS: DNA sequencing and restriction enzyme digestion proved that HER-2 gene was correctly connected. SDS-PAGE and Western blotting analysis showed that the protein was expressed in conclusion body in E. coli DH5α and bound to Herceptin. We purified the recombinant protein by Ni-NAT affinity chromatography and obtained soluble human HER-2 extracellular domain. Immune response was effectively induced by rhHER-2. CONCLUSION: HER-2 protein vaccine has been successfully prepared and effective immune response has been induced.
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万方数据
维普
