DDAB/PLA纳米疫苗佐剂放大工艺研究及初步安全性评估
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摘要
采用减压蒸发结合切向流超滤法放大制备了1种用作疫苗佐剂的双十八烷基二甲基溴化铵(DDAB)修饰的聚乳酸(PLA)纳米球(DDAB/PLA),并对制备的纳米球进行表征,包括表面形貌、粒径及粒度分布、ζ电位、抗原吸附率、收率、稳定性。以健康雌性Balb/c小鼠为实验对象,进行纳米球初步安全性评价。结果表明,制备所得纳米球粒径为(150.3±10.4)nm;ζ电位为(54.0±4.0)mV;产率为(96.3±5.2)%;对鸡卵清白蛋白(OVA)吸附率为(85.4±0.4)%。扫描电镜结果显示所制备的纳米球呈圆整球形,且分布均一。该疫苗佐剂体外稳定性较好,在超过正常注射剂量100倍的情况下,纳米球对小鼠具有良好的安全性。
The dimethyldioctadecylammonium bromide(DDAB) modified polylactide(PLA) nanoparticles used as vaccine adjuvant were prepared using vacuum evaporation combined with tangential flow ultrafiltration in scaleup level. The surface morphology, particle size and size distribution, ζ potential, ovalbumin(OVA) antigen loading efficiency, yield and stability were also characterized. The materials were evaporated for 6 h in vacuum condition followed by tangential flow ultrafiltration to obtain the optimal products. The results showed that the average particle size,ζ potential, yield and OVA antigen loading efficiency of the nanoparticles with spherical shapes were(150.3±10.4)nm,(54.0±4.0)mV,(96.3±5.2)% and(85.4±0.4)%, respectively. These nanoparticles kept stable after storing at 4 ℃ for 15 d. The preliminary safety evaluation of the nanoparticles was carried out with healthy female Balb/c mice as experimental subjects. The results of HE staining slice showed that DDAB/PLA nanoparticles had no obvious side effects on the main organs of mice at a dose exceeding 100 times of the normal injection dose, indicating that the nanoparticles showed good safety.
The dimethyldioctadecylammonium bromide(DDAB) modified polylactide(PLA) nanoparticles used as vaccine adjuvant were prepared using vacuum evaporation combined with tangential flow ultrafiltration in scaleup level. The surface morphology, particle size and size distribution, ζ potential, ovalbumin(OVA) antigen loading efficiency, yield and stability were also characterized. The materials were evaporated for 6 h in vacuum condition followed by tangential flow ultrafiltration to obtain the optimal products. The results showed that the average particle size,ζ potential, yield and OVA antigen loading efficiency of the nanoparticles with spherical shapes were(150.3±10.4)nm,(54.0±4.0)mV,(96.3±5.2)% and(85.4±0.4)%, respectively. These nanoparticles kept stable after storing at 4 ℃ for 15 d. The preliminary safety evaluation of the nanoparticles was carried out with healthy female Balb/c mice as experimental subjects. The results of HE staining slice showed that DDAB/PLA nanoparticles had no obvious side effects on the main organs of mice at a dose exceeding 100 times of the normal injection dose, indicating that the nanoparticles showed good safety.
引文
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[5]堵祖荫.化工工艺开发放大的方法[J].化工与医药工程,2017,38(3):1-3.
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[7]邱越佳,孟丹,杨婷媛,等.口服载胰岛素PLGA纳米球的制备及初步药效学评价[J].中国医药工业杂志,2018,49(5):614-621.
[8]国家食品药品监督管理局药品审评中心.GPT2-1.预防用生物制品临床前安全性评价技术审评一般原则[S].2005.
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