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.

胶质母细胞瘤是成年人中最常见的原发性恶性脑肿瘤。目前，外科手术切除肿瘤块，随后进行辅助放疗和化学疗法是多种形式的胶质母细胞瘤的标准治疗方法，但到目前为止仍不是有效的治疗方法。因此，非常需要开发一种疫苗，作为针对胶质母细胞瘤多种形式的预防或治疗目的的安全有效策略。本研究旨在设计针对多形性胶质母细胞瘤的多域疫苗。一个在硅片该方法用于选择最有效的蛋白质结构域，以诱导宿主针对多形性胶质母细胞瘤的B细胞和T细胞免疫应答。发现IL-13Rα-2（氨基酸位置27–144），TNC（氨基酸位置1900–2100）和PTPRZ-1（氨基酸位置731–884）具有有效的诱导免疫反应。因此，我们认为它们与链接器A（EAAAK）₃融合A构建多域重组疫苗。对设计的重组疫苗构建体进行了免疫信息学分析，以评估其功效。尽管设计的重组疫苗构建体没有显示过敏原特性，但其抗原性估计为0.78。表征了重组疫苗构建体的理化性质，并揭示了候选疫苗的效力。然后使用生物信息学工具预测其二级和三级结构，mRNA结构，分子对接和免疫模拟。接下来，合成设计的重组疫苗构建体，并将其克隆到pET28a载体中并在大肠杆菌中表达BL21。此外，利用圆二色谱法研究了重组疫苗构建体的二级结构变化。重组疫苗构建体表达的验证评估结果表明，计算机分析是相对准确的，蛋白质二级结构发生了相对变化。在我们的未来计划中，应通过进一步的体外和体内实验研究来验证通过计算机工具确认的候选疫苗。