Department of Biotechnological Development, Hygiene Institute, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay.
Cellular and Molecular Biology Laboratory, Center for Biotechnology-CBiotec, Federal University of Paraíba, João Pessoa, Paraíba, Brazil.
Methods Mol Biol. 2022;2469:183-191. doi: 10.1007/978-1-0716-2185-1_15.
Adjuvants are essential components of subunit, recombinant, nonreplicating and killed vaccines, as they are substances that boost, shape, and/or enhance the immune response triggered by vaccination. Saponins obtained from the Chilean Q. saponaria tree are used as vaccine adjuvants in commercial vaccines, although they are scarce and difficult to obtain. In addition, tree felling is needed during its extraction, which has ecological impact. Q. brasiliensis leaf-extracted saponins arise as a more sustainable alternative, although its use is still limited to preclinical studies. Despite the remarkable immunostimulating properties of saponins, they are toxic to mammalian cells, due to their intrinsic characteristics. For these reasons they are mostly used in veterinary vaccines, although recently the Q. saponaria purified saponin QS-21 has been included in adjuvant systems for human vaccines, such as Mosquirix and Shingrix (GSK). In order to abrogate the toxicity of the saponins fractions, they can be formulated as immunostimulating complexes (ISCOMs). ISCOM-matrices are cage-like nanoparticles of approximately 40 nm, formulated combining saponins and lipids, without antigen, and are great adjuvants able to promote Th1-biased immune responses in a safe manner. Herein we describe how to formulate ISCOM-matrices nanoparticles using Q. brasiliensis purified saponin fractions (IMXQB) by the dialysis method. In addition, we indicate how to verify the appropriate size and homogeneity of the formulated nanoparticles.
佐剂是亚单位疫苗、重组疫苗、非复制疫苗和灭活疫苗的重要组成部分,因为它们是增强、塑造和/或增强疫苗接种引发的免疫反应的物质。从智利 Q.saponaria 树中获得的皂苷被用作商业疫苗中的疫苗佐剂,尽管它们稀缺且难以获得。此外,在提取过程中需要砍伐树木,这对生态有影响。从 Q.brailiensis 叶中提取的皂苷作为一种更可持续的替代品出现,尽管其用途仍限于临床前研究。尽管皂苷具有显著的免疫刺激特性,但由于其固有特性,它们对哺乳动物细胞有毒。出于这些原因,它们主要用于兽医疫苗,尽管最近 Q.saponaria 纯化的皂苷 QS-21 已被纳入人类疫苗的佐剂系统,如 Mosquirix 和 Shingrix(GSK)。为了消除皂苷部分的毒性,可以将其制成免疫刺激复合物(ISCOMs)。ISCOM-基质是大约 40nm 的笼状纳米颗粒,通过组合皂苷和脂质,不添加抗原来配方,是能够以安全的方式促进 Th1 偏向性免疫反应的良好佐剂。本文描述了如何通过透析法使用从 Q.brailiensis 纯化的皂苷(IMXQB)来配方 ISCOM-基质纳米颗粒。此外,我们还指出了如何验证配方纳米颗粒的适当大小和均一性。