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基于天然油脂的均一、稳定的乳液制备及其作为疫苗佐剂的研究
陈增辉
学位类型硕士
导师苏志国 ; 张松平
2016-07
学位授予单位中国科学院研究生院
学位授予地点北京
学位专业生物化工
关键词油脂 W/o乳液 O/w乳液 稳定性 佐剂
摘要

油乳佐剂是常用的一种佐剂类型,在人类和动物疫苗领域均有广泛应用。作为疫苗抗原的递送载体,乳液应具有良好的稳定性和生物安全性。传统的乳液佐剂大多数基于矿物质油,难以生物降解,副作用严重。本课题以植物来源的油脂为油相制备稳定的、具有良好生物相容性的乳液,并将其应用于重要的兽用疫苗口蹄疫灭活病毒(foot-and-mouth disease virus, FMDV)疫苗和模式抗原卵清蛋白(ovalbumin,OVA)的佐剂,对其佐剂效果进行评价。本文取得的主要结果如下:(1)以肉豆蔻酸异丙酯为油相,聚甘油醇蓖麻醇酯(PGPR)为表面活性剂制备w/o乳液。通过在水相中添加亲水性表面活性剂Tween 80,可以显著提高乳液的稳定性。其主要稳定性机理包括降低乳液粒径、提高粒径分布的均一性、降低油水界面张力、增加乳液粘度等。当油水质量比为2:1、PGPR和Tween 80在油、水相中的质量比分别为5%的条件下,可以制备出平均粒径为130 nm,稳定的w/o纳米乳液;(2)以肉豆蔻酸异丙酯为油相,Tween 20为水相表面活性剂制备o/w乳液。通过在水相中加入黄原胶,可以显著提高乳液的稳定性。在油水质量比为1:1、Tween 20浓度为5%、黄原胶浓度为0.08%条件下,可以制备出粒径为3.5 mm的稳定、均一的o/w乳液;(3)将上述制备的w/o纳米乳液和o/w微米乳液包埋FMDV和模式抗原卵清蛋白(ovalbumin),考察其作为佐剂的性能。结果表明与商品化的Montanide ISA206乳液佐剂相比,所制备的乳液为佐剂均可以显著提高抗原的体液免疫和细胞免疫水平,尤其是在激发细胞免疫水平方面,甚至显著优于ISA206佐剂。本论文的研究结果,为发展新一代可替代基于矿物油佐剂的、具有更好生物安全性、更有效增强体液免疫和细胞免疫水平的乳液佐剂,提供新的思路。

其他摘要

Adjuvant based on oil emulsions is a common type of all adjuvants, widely used in the field of human and animal vaccines. It is essential for emulsion with good stability and biological safety as an antigen delivery system. Traditional emulsion adjuvants are mostly based on mineral oil and hard to be degraded, resulting in side effects. This thesis focused on the oils and surfactants from vegetables, and prepared emulsions with them to formulate stable and bio-compatible emulsions. Foot-and-mouth disease virus (FMDV) and ovalbumin (OVA) were used to test the potential adjuvant effects of these emulsions. The main work done in this thesis is as follow:(1) Stable water-in-oil (w/o) emulsion was prepared with isopropyl myristate as oil phase and polyglycerol polyricinoleate as emulsifier. Additionally, Tween 80 was added to aqueous phase before emulsification to improve the stability of emulsion stability. The effect of Tween 80 on stable w/o emulsion resulting from decreasing the particle size, narrowing size distribution, decreasing interfacial tension between oil and water, and increasing emulsion viscosity. The optimum condition of w/o emulsion preparation system was set with ratio of oil and water 2:1, 5% PGPR in oil phase and 5% Tween 80 in aqueous phase, thereafter, the stable w/o emulsion was obtained with particle size of 130 nm diameter.(2) Stable oil-in-water (o/w) emulsion was prepared with isopropyl myristate as oil phase and Tween 20 as emulsifier. Xanthan gum was added in aqueous phase as the stabilizer. When the conditions for emulsification was set with oil / water ratio 1:1, 5% Tween 20 and 0.08% xanthan gum in aqueous phase, the stable o/w emulsion with particle size of 3.5 mm was obtained.(3) The stable w/o and o/w emulsions above equipped with FMDV and a model antigen OVA were used to evaluate the potential adjuvant effects. Both w/o and o/w emulsions could enhance humoral and cellular immune compared to Montanide ISA206, especially in cellular immune response.The emulsions developed in this thesis could provide some novel ideas for preparation of emulsion adjuvants, which could be an alternative to the mineral oil adjuvants and have significant humoral and cellular immune response with better biological safety. 

语种中文
文献类型学位论文
条目标识符http://ir.ipe.ac.cn/handle/122111/22865
专题研究所(批量导入)
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陈增辉. 基于天然油脂的均一、稳定的乳液制备及其作为疫苗佐剂的研究[D]. 北京. 中国科学院研究生院,2016.
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