Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran.
Department of Pathobiology, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Arch Razi Inst. 2022 Feb 28;77(1):37-44. doi: 10.22092/ari.2021.353322.1596. eCollection 2022 Feb.
Foot-and-mouth disease (FMD) is an acute and highly contagious disease in livestock, such as cattle, sheep, and pigs, leading to a lot of economic losses. The current FMD vaccines formulated by inactivated whole-virus and adjuvant successfully reduce disease outbreaks in many regions of the world. Immunological studies on FMD viruses revealed that the dominant epitope in arising neutral antibody response is amino acid residues constructing the G-H loop, constituting a surface loop of the structural protein, termed VP1. Liposomes as one of the most well-known vehicles are considered an important carrier in vaccine development, and their function is used to encapsulate purified VP1 protein based on their size, charge, and lipid content. Accordingly, the VP1 protein was isolated from the FMD virus. This study aimed to compare four methods of VP1 protein encapsulation in the liposome and the extruding effect, as follows: 1) VP1 protein was dissolved in dimethyl sulfoxide and added to the lipid film hydrated by ethanol, 2) the lipid film was hydrated by VP1 protein with 7M urea, 3) the lipid film was hydrated by VP1 protein and freeze-thawed, and 4) the lipid film was hydrated by VP1 protein. The highest encapsulation efficiency was 91% in the second method which purified protein-containing urea. The VP1 protein in the prepared liposome (1, 2-dimyristoyl-sn-glycero-3-phosphocholine: 1, 2-dimyristoyl-sn-glycero-3-phosphocholine: cholesterol) released more than 90% of protein content after 240 h.
口蹄疫(FMD)是一种急性、高传染性的牲畜疾病,如牛、羊、猪等,导致了大量的经济损失。目前,使用灭活全病毒和佐剂制成的 FMD 疫苗在世界许多地区成功地减少了疾病的爆发。对口蹄疫病毒的免疫研究表明,在产生中和抗体的反应中,优势表位是构成结构蛋白 G-H 环的氨基酸残基,称为 VP1。脂质体作为最著名的载体之一,被认为是疫苗开发的重要载体,其功能是根据其大小、电荷和脂质含量,将纯化的 VP1 蛋白包裹在脂质体中。因此,从 FMD 病毒中分离出 VP1 蛋白。本研究旨在比较 VP1 蛋白包封在脂质体中的四种方法及其挤出效果,如下:1)VP1 蛋白溶解在二甲基亚砜中,加入乙醇水合的脂质膜;2)脂质膜由含 7M 尿素的 VP1 蛋白水合;3)脂质膜由 VP1 蛋白冷冻解冻水合;4)脂质膜由 VP1 蛋白水合。第二种方法的包封效率最高,为 91%,该方法中纯化的含脲蛋白。制备的脂质体(1,2-二肉豆蔻酰基-sn-甘油-3-磷酸胆碱:1,2-二肉豆蔻酰基-sn-甘油-3-磷酸胆碱:胆固醇)中的 VP1 蛋白在 240 小时后释放出超过 90%的蛋白含量。