基于Box-Behnken试验优化的枸杞子多糖油乳佐剂配方的建立  

Optimization of Lycium bararum polysaccharides adjuvant formulation and its preliminary characterization using Box-Behnken experiment

作　　者：杨坤 孙世琪[2] 石晓妮[2] 董虎[2] 赵凯 郭慧琛[2,3] 魏凡华 YANG Kun;SUN Shi-qi;SHI Xiao-ni;DONG Hu;ZHAO Kai;GUO Hui-chen;WEI Fan-hua(College of Agriculture,Ningxia Universty,Yinchuan 750021,China;State Key Laboratory of Veterinary Etiological Biology/National Foot-and-Mouth Disease Reference Laboratory/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory,Yunnan Animal Science and Veterinary Institute,Kunming 650224,China;Institute of Nanobiomaterials and Immunology,School of Life Science,Taizhou University,Taizhou 318000,China)

机构地区：[1]宁夏大学农学院,宁夏银川750021 [2]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,国家口蹄疫参考实验室,甘肃兰州730046 [3]云南热带亚热带动物病毒病实验室云南省畜牧兽医科学院,云南昆明650224 [4]台州学院纳米生物材料与免疫学研究所生命科学学院,浙江台州318000

出　　处：《中国兽医科学》2022年第9期1122-1129,共8页Chinese Veterinary Science

基　　金：国家自然科学基金项目(3207284,31873023,32072859,32002272);国家重点研发计划项目(2021YFD1800300);科技人才与平台计划项目(202205AF150007)。

摘　　要：为优化枸杞子多糖油乳佐剂(LBPOA)的最佳配方,使用三因素二水平Box-Behnken试验设计,以枸杞子多糖为药物模型,磁悬搅拌法制备LBPOA。采用响应曲面法对影响乳液成乳能力的主要因素进行考察,枸杞子多糖油乳佐剂承载枸杞子多糖的质量作为响应值,用二项式数学模型对配方进行优化。以口蹄疫病毒样颗粒(foot-and-mouth disease virus-like particles,FMD-VLPs)为抗原模型,考察LBPOA的生物相容性和对疫苗生物活性的影响。结果,枸杞子多糖油乳佐剂的最佳成乳条件为:33.905%的EL-35,LBP的浓度为9.996 mg/m L,37.55%的无水乙醇,最佳载水量为9.33%。LBPOA在10000 r/min离心10 min,粒径、电位、PDI均无显著变化。于4℃、25℃和37℃条件下将其放置7 d,粒径、PDI无较大波动。优化后的枸杞子多糖油乳佐剂与FMD-VLPs乳化免疫小鼠,对小鼠主要脏器无明显损伤。37℃条件下,LBPOA包封FMDVLPs,抗原活性没有大幅度下降。上述结果表明,Box-Behnken试验设计能够有效优化LBPOA配方,可作为其他疫苗佐剂及复合物配方的分析模型及有效手段;LBPOA具备良好的稳定性、生物相容性,能够维持蛋白质抗原的生物活性。The formulation of Lycium barbarum Polysaccharide(LBP)oil emulsion adjuvant(LBPOA)was optimized by the Box-Behnken experimental design.Moreover,the preparation of LBPOA by magnetic stirring using LBP as a drug model.Response surface methodology was applied to examine the main influential factors of emulsion formation.The response value was defined by the mass of LBPOA-loaded LBP solution and the prescription was optimized using a binomial mathematical model.Foot-and-mouth disease virus-like particles were used as a vaccine model to investigate the protective effect of LBPOA on vaccine.The results of the research show that the appropriate formulation of LBPOA was composed of 33.905% EL-35,37.55% anhydrous ethanol,and 9.996 mg/m L of concentration of LBP solution,which has a most suitable loading of 9.33%.LBPOA was centrifuged at 10000 r/min for 10 min,with no significant change in particle size,potential,or PDI,which was stored for 7 days at different temperatures(37℃,4℃,and 25℃)with no significant change in particle size or PDI.Animal experiments were conducted to evaluate the effect of the LBPOA in mice.The experimental results showed that LBPOA caused no damage to the internal organs of mice.LBPOA loaded with FMD-VLPs was placed at 37℃ for 7 days,it caused no significant decrease in the content of FMD-VLPs.In conclusion,Box-Behnken experimental design is effective in optimizing LBPOA formulations,which is an analytical model and an effective method for adjuvant and compound formulations.In addition,LBPOA has good stability,biocompatibility,and maintenance of the biological activity of the vaccine.

关 键 词：Box-Behnken试验 枸杞子多糖 油乳佐剂 口蹄疫病毒样颗粒 

分 类 号：S853.733[农业科学—临床兽医学]