Development of low-cost cage-like particles to formulate veterinary vaccines

卵清蛋白 蛋黄 明矾 佐剂 薄层色谱法 色谱法 抗体 免疫系统 生物 化学 食品科学 免疫学 有机化学
作者
Giuliana Lupi,Florencia X. Santiago Valtierra,Gabriel Cabrera,Roque Spinelli,Álvaro S. Siano,Verónica González,Antonio Osuna,Gerardo M. Oresti,Iván Marcipar
出处
期刊:Veterinary Immunology and Immunopathology [Elsevier]
卷期号:251: 110460-110460
标识
DOI:10.1016/j.vetimm.2022.110460
摘要

Low-cost adjuvants are urgently needed for the development of veterinary vaccines able to trigger strong immune responses. In this work, we describe a method to obtain a low-cost cage-like particles (ISCOMATRIX-like) adjuvant useful to formulate veterinary vaccines candidates. The main components to form the particles are lipids and saponins, which were obtained from egg yolk by ethanolic extraction and by dialyzing a non-refined saponins extract, respectively. Lipids were fully characterized by thin layer chromatography (TLC) and gas-chromatography (GC) and enzymatic methods, and saponins were characterized by TLC, HPLC and MALDI-TOF. Cage-like particles were prepared with these components or with commercial inputs. Both particles and the traditional Alum used in veterinary vaccines were compared by immunizing mice with Ovalbumin (OVA) formulated with these adjuvants and assessing IgG1, IgG2a anti OVA antibodies and specific Delayed-type Hypersensitivity (DTH). In the yolk extract, a mixture of phospholipids, cholesterol and minor components of the extract (e.g. lyso-phospholipids) with suitable proportions to generate cage-like particles was obtained. Also, semi-purified saponins with similar features to those of the QuilA® were obtained. Cage-like particles prepared with these components have 40-50 nm and triggers similar levels of Anti-OVA IgG1 and DTH than with commercial inputs but higher specific-IgG2a. Both adjuvants largely increased the levels of IgG1, IgG2a and DTH in relation to the formulation with Alum. The methods described to extract lipids from egg yolk and saponins from non-refined extract allowed us to obtain an inexpensive and highly effective adjuvant.
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