Monkeypox, a viral zoonotic disease resembling smallpox, has emerged as a significant national epidemic primarily in Africa. Nevertheless, the recent global dissemination of this pathogen has engendered apprehension regarding its capacity to metamorphose into a sweeping pandemic. To effectively combat this menace, a multi-epitope vaccine has been meticulously engineered with the specific aim of targeting the cell envelope protein of Monkeypox virus (MPXV), thereby stimulating a potent immunological response while mitigating untoward effects. This new vaccine uses T-cell and B-cell epitopes from a highly antigenic, non-allergenic, non-toxic, conserved, and non-homologous A30L protein to provide protection against the virus. In order to ascertain the vaccine design with the utmost efficacy, protein–protein docking methodologies were employed to anticipate the intricate interactions with Toll-like receptors (TLR) 2, 3, 4, 6, and 8. This meticulous approach led the researchers to discern an optimal vaccine architecture, bolstered by affirmative prognostications derived from both molecular dynamics (MD) simulations and immune simulations. The current research findings indicate that the peptides ATHAAFEYSK, FFIVVATAAV, and MNSLSIFFV exhibited antigenic properties and were determined to be non-allergenic and non-toxic. Through the utilization of codon optimization and in-silico cloning techniques, our investigation revealed that the prospective vaccine exhibited a remarkable expression level within Escherichia coli. Moreover, upon conducting immune simulations, we observed the induction of a robust immune response characterized by elevated levels of both B-cell and T-cell mediated immunity. Moreover, as the initial prediction with in-silico techniques has yielded promising results these epitope-based vaccines can be recommended to in vitro and in silico studies to validate their immunogenic properties.

猴痘是一种类似天花的病毒性人畜共患疾病，已成为主要在非洲的重要全国性流行病。然而，这种病原体最近在全球范围内的传播引起了人们对其演变成一场大流行的能力的担忧。为了有效对抗这一威胁，我们精心设计了一种多表位疫苗，其具体目标是针对猴痘病毒（MPXV）的细胞包膜蛋白，从而刺激有效的免疫反应，同时减轻不良影响。这种新疫苗使用来自高抗原性、非过敏性、无毒、保守且非同源的 A30L 蛋白的 T 细胞和 B 细胞表位来提供针对病毒的保护。为了确定疫苗设计的最大功效，采用蛋白质-蛋白质对接方法来预测与 Toll 样受体 (TLR) 2、3、4、6 和 8 之间复杂的相互作用。这种细致的方法使研究人员在分子动力学（MD）模拟和免疫模拟得出的肯定预测的支持下，识别最佳的疫苗架构。目前的研究结果表明，肽ATHAAFEYSK、FFIVVATAAV和MNSLSIFFV表现出抗原特性，并被确定为非过敏性和无毒的。通过利用密码子优化和计算机克隆技术，我们的研究表明，该预期疫苗在大肠杆菌中表现出显着的表达水平。此外，在进行免疫模拟时，我们观察到诱导了强大的免疫反应，其特征是 B 细胞和 T 细胞介导的免疫水平升高。此外，由于计算机技术的初步预测已经产生了有希望的结果，因此可以推荐这些基于表位的疫苗进行体外和计算机研究，以验证其免疫原性。