Evaluation of Oral and Injection Immunization with Chitosan Nanoparticles ‎Containing HER-۲ antigene

Aims: Breast cancer is the most common cancer among women around the world and in Iran and ۲۲% of all cancers in women is included. About ۳۰% of cases with breast cancers are due to the proliferation or excessive expression of HER-۲ protein. Chitosan is an environmentally friendly combination, due to its minimal systemic toxicity of peptide or drug delivery, the application is considered. The aims of this study were to manufactur chitosan nanoparticle containing HER-۲ recombinant protein, evaluation of the properties and effect of its oral and injection immunization on spleen lymphocytes. Materials and Methods: In this experimental study, pET-۲۸a vector containing HER-۲ gene was evaluated using PCR by universal primers. Expression of protein in E.coli was induced with IPTG. The recombinant protein was purified using affinity chromatography and evaluated by Western Blotting analysis. Chitosan nanoparticles containing recombinant protein were prepared by inonic gelation method and their size were characterized by DLS. Mice were immunized with nanoparticles and antibody titers were determined by ELISA. The response of lymphocytes in exposure to nanoparticles was evaluated by MTT assay. One way ANOVA, Duncan test and SPSS ۲۲ software were used for statistical analysis. Findings: The protein with a molecular weight of ۱۸ kDa was confirmed. Yield of protein was ۱۲mgL-۱. Encapsulation efficiency of recombinant protein in nanoparticles was ۷۰%. The average particle size was ۲۰۵.۲nm. Immunization of mice induced mucosal and humoral immune response. Conclusion: Encapsulation efficiency of recombinant protein in nanoparticles is ۷۰%. The average particle size is ۲۰۵.۲nm. Immunization of mice induces mucosal and humoral immune response HER-۲ recombinant protein, due to the cellular and humoral immune response, is a candidate suitable for cancer diagnostic or therapeutic purposes.

کلیدواژه ها:

Breast Neoplasms ، Chitosan ، HER-۲ Protein ، ‎Nanoparticle ، بدخیمی های پستان ، کیتوزان ، پروتئین HER-۲‎ ، نانوذره