Department of Eukaryote Gene Engineering, Institute of Biotechnology, Life Sciences Center, Vilnius University, Sauletekio al. 7, Vilnius, Lithuania.
Department of Eukaryote Gene Engineering, Institute of Biotechnology, Life Sciences Center, Vilnius University, Sauletekio al. 7, Vilnius, Lithuania.
Virus Res. 2021 Jan 15;292:198236. doi: 10.1016/j.virusres.2020.198236. Epub 2020 Nov 23.
Human parvovirus 4 (PARV4) is a novel tetraparvovirus that was isolated from intravenous drug users in 2005. Recombinant PARV4 capsid protein VP2 can form stable virus-like particles (VLPs) in yeast. These VLPs could act as antigen carriers during vaccine development. Therefore, the information about PARV4 VP2 VLP antigenic sites could advance further research in this area. In this work, human parvovirus 4 VLPs obtained from yeast were used to generate monoclonal antibodies (mAbs) in mice. Epitope mapping of the obtained mAbs showed at least three distinct antigenic sites of the VP2 protein. On top of that, molecular cloning was used to replace PARV4 VP2 antigenic sites with heterologous peptides. The chimeric PARV4 VLPs bearing polyhistidine inserts obtained from yeast were observed using electron microscopy while polyhistidine-specific antibodies detected heterologous peptides of the chimeric VP2 proteins.
人细小病毒 4 型(PARV4)是一种新型的四联体细小病毒,于 2005 年从静脉吸毒者中分离得到。重组 PARV4 衣壳蛋白 VP2 可在酵母中形成稳定的病毒样颗粒(VLPs)。这些 VLPs 可在疫苗开发过程中作为抗原载体。因此,PARV4 VP2 VLP 抗原表位的信息可以推进该领域的进一步研究。在这项工作中,从酵母中获得的人细小病毒 4 型 VLPs 被用于在小鼠中产生单克隆抗体(mAbs)。获得的 mAbs 的表位作图显示,VP2 蛋白至少有三个不同的抗原表位。除此之外,还使用分子克隆技术将 PARV4 VP2 抗原表位替换为异源肽。从酵母中获得的带有组氨酸插入的嵌合 PARV4 VLPs 通过电子显微镜观察,而组氨酸特异性抗体检测嵌合 VP2 蛋白的异源肽。