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Updated: Nov 24, 2025
Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells
Published on: November 28, 2015
Immuno-informatics analysis and expression of a novel multi-domain antigen as a vaccine candidate against
Mahmoud Gharbavi1, Hossein Danafar2, Jafar Amani3
1Department of Pharmaceutical Biomaterials, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran; Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
Abstract:
Glioblastoma multiform is the most common of primary malignant brain tumors in adults. Currently, surgical resection of the tumor mass, followed by adjuvant radiotherapy and chemotherapy are standard treatments for glioblastoma multiform but so far are not effective treatments. Thus, the development of a vaccine, as a safe and efficient strategy for prophylactic or therapeutic purposes against glioblastoma multiform is very necessary. The present study aimed to design the multi-domain vaccine for glioblastoma multiform. An in silico approach was used to select the most potent domains of proteins to induce the host's B- and T-cell immune response against glioblastoma multiform. IL-13Rα-2 (amino acid positions 27-144), TNC (amino acid positions 1900-2100), and PTPRZ-1(amino acid positions 731-884) were found to have potent inducible immune responses. So, we considered them for fusing with a linker A(EAAAK)3A to construct the multi-domain recombinant vaccine. The immuno-informatics analysis of the designed recombinant vaccine construct was performed to evaluate its efficacy. Although the designed recombinant vaccine construct did not show allergen property, its antigenicity was estimated at 0.78. The Physico-chemical properties of the recombinant vaccine construct were characterized and revealed the potency of the vaccine candidate. Then its secondary and tertiary structures, mRNA structure, molecular docking, and immune simulation were predicted using bioinformatics tools. Next, the designed recombinant vaccine construct was synthesized, and cloned into the pET28a vector and expressed in E. coli BL21. Besides, the circular dichroism spectroscopy was utilized for the investigation of the secondary structure changes of the recombinant vaccine construct. The results of the verification assessment of the recombinant vaccine construct expression indicated that in silico analysis was relatively accurate, and relatively change occurred on the protein secondary structure. In our future plan, the vaccine candidate that was confirmed by in silico tools should be validated by further in vitro and in vivo experimental studies.
Insights
This study designed a novel multi-domain vaccine for glioblastoma multiform using in silico methods. The vaccine shows promise for future glioblastoma treatment by eliciting immune responses.
Area of Science:
- Oncology
- Immunology
- Bioinformatics
Background:
- Glioblastoma multiform is a primary malignant brain tumor with limited treatment options.
- Current treatments like surgery, radiotherapy, and chemotherapy are largely ineffective.
- There is a critical need for novel prophylactic or therapeutic strategies, such as vaccines, against glioblastoma.
Purpose of the Study:
- To design a multi-domain recombinant vaccine for glioblastoma multiform.
- To identify potent protein domains that can induce B- and T-cell immune responses against glioblastoma.
- To evaluate the designed vaccine construct's efficacy using in silico and experimental methods.
Main Methods:
- In silico selection of immunogenic protein domains: IL-13Rα-2, TNC, and PTPRZ-1.
- Construction of a multi-domain recombinant vaccine by fusing selected domains with a linker.
- In silico analysis including immuno-informatics, physico-chemical properties, structural prediction, molecular docking, and immune simulation.
- Experimental validation including synthesis, cloning, expression in E. coli, and circular dichroism spectroscopy.
Main Results:
- Identified IL-13Rα-2, TNC, and PTPRZ-1 domains as potent inducers of immune responses.
- Designed a multi-domain recombinant vaccine construct with estimated antigenicity of 0.78 and no allergenicity.
- Confirmed the accuracy of in silico predictions through experimental expression and secondary structure analysis.
- Observed minor changes in the protein's secondary structure upon expression.
Conclusions:
- The designed multi-domain recombinant vaccine construct shows potential for glioblastoma treatment.
- In silico approaches are effective in designing and predicting the properties of vaccine candidates.
- Further in vitro and in vivo studies are required to validate the efficacy of this glioblastoma vaccine candidate.
