Volume 21, Issue 2 (2018)
mjms 2018, 21(2): 85-94 |
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Shafighian F, Nazarian S, Amani J, Fahimi H. Evaluation of Oral and Injection Immunization with Chitosan Nanoparticles Containing HER-2 antigene. mjms 2018; 21 (2) :85-94
URL: http://mjms.modares.ac.ir/article-30-1656-en.html
URL: http://mjms.modares.ac.ir/article-30-1656-en.html
Evaluation of Oral and Injection Immunization with Chitosan Nanoparticles Containing HER-2 antigene
1- Pharmaceutical Sciences Research Center, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
2- Biology Department, Basic Sciences Faculty, Imam Hossain University, Tehran, Iran ,Nazarian56@gmail.com
3- Applied Microbiology Research Center, System biology and poisonings institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
4- Molecular and Cellular Sciences Department, Advanced Sciences and Technology Faculty, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
2- Biology Department, Basic Sciences Faculty, Imam Hossain University, Tehran, Iran ,
3- Applied Microbiology Research Center, System biology and poisonings institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
4- Molecular and Cellular Sciences Department, Advanced Sciences and Technology Faculty, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
Abstract: (9315 Views)
Aims: Breast cancer is the most common cancer among women around the world and in Iran and 22% of all cancers in women is included.
About 30% of cases with breast cancers are due to the proliferation or excessive expression of HER-2 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-2 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-28a vector containing HER-2 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 22 software were used for statistical analysis.
Findings: The protein with a molecular weight of 18 kDa was confirmed. Yield of protein was 12mgL-1. Encapsulation efficiency of recombinant protein in nanoparticles was 70%. The average particle size was 205.2nm. Immunization of mice induced mucosal and humoral immune response.
Conclusion: Encapsulation efficiency of recombinant protein in nanoparticles is 70%. The average particle size is 205.2nm. Immunization of mice induces mucosal and humoral immune response HER-2 recombinant protein, due to the cellular and humoral immune response, is a candidate suitable for cancer diagnostic or therapeutic purposes.
Article Type: Original Manuscipt |
Subject:
Biochemistry
Received: 2017/10/28 | Accepted: 2018/02/12
Received: 2017/10/28 | Accepted: 2018/02/12
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