Iranian Journal of Medical Sciences

Document Type : Original Article(s)


Department of Medical Biotechnology, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran



Background: Hepatocellular carcinoma (HCC) is one of the prevalent cancers in the world with a high recurrence rate. In recent years, different researches have focused on designing efficient multi-epitope peptide vaccines against HCC. In designing these vaccines, over-expressed antigens in HCC patients, such as α- fetoprotein (AFP) and Glypican-3 (GPC-3), have been considered. In our previous study, a multi-epitope peptide vaccine for HCC was designed by in silico methods. The designed vaccine construct included the AFP, GPC-3, and Aspartyl-β-hydroxylase (ASPH) as CytoLoxic T cell Lymphocytes (CTL), one epitope from Tetanus Toxin Fragment C (TTFrC) as Helper T cell Lymphocytes (HTL), and a segment of microbial heat shock protein (HSP70) peptide 407-426 as an adjuvant. All the mentioned parts were connected by appropriate linkers. The aim of this study is the production of the designed vaccine.
Methods: This research is experimental and was carried out in Fasa, Iran, in 2017. The designed vaccine construct was transformed to the E.coli BL21 (DE3) strain and expressed in different Isopropyl β-D-1-thiogalactopyranoside (IPTG) concentrations (0.6 and 1 mM), times (4, 6, 8, 16 hours), and temperatures (25 and 37 °C). Then, the expressed protein was analyzed by Sodium Dodecyl Sulfate-polyacrylamide (SDS) gel and the Western blot method.
Results: The best conditions for protein expression were obtained in the Super Optimal Broth (SOB) medium at 37 °C after the induction of expression by 1 mM IPTG for six hour.
Conclusion: The recombinant HCC vaccine was produced with a proper concentration.


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