Thermal Treatment of a Novel Saponin-Cholesterol Nanoparticle Vaccine Adjuvant Named NanoQuil F70 Secures a Uniform Morphology and Size Distribution

Ke-fei Hu, L. Duroux, E. B. Lindblad
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Abstract

Quil-A and cholesterol can, under certain conditions, form nanoparticles, but the conditions for the formation of a homogeneous population of a particular kind of particles with the same morphology and size have remained elusive. However, a well-defined uniformity, as well as a high degree of batch-to-batch consistency, are prerequisites for adjuvant formulations to be used for practical vaccines. To accomplish the above stated tasks in the present study, we demonstrated that Quil-A and cholesterol form well-defined worm-like stable particles with a mean diameter of around 40 nm by dynamic light scattering (DLS) under carefully controlled thermodynamical conditions with little or no Quil-A degradation. The nanoparticles thus prepared possessed a significantly reduced hemolytic effect in comparison to unformulated free Quil-A. However, when the same conditions were applied to QS-21/cholesterol nanoparticle formation the morphology achieved was coil spring-like particles with a mean diameter of around 70 nm. This discrepancy in particle morphology and size was attributed to the differences in hydrophobicity of the Quil-A, being a heterogenic fraction of acylated as well as deacylated saponins, and the QS-21. With the process optimizations leading to a standardized particle size distribution and nanoparticle morphology presented here, NanoQuil F70 holds the potential as a well-tolerated vaccine adjuvant for veterinary use raising a Th1/Th2 balanced immune response.
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新型皂苷-胆固醇纳米颗粒疫苗佐剂NanoQuil F70的热处理确保了其均匀的形态和大小分布
quila和胆固醇在一定条件下可以形成纳米颗粒,但形成具有相同形态和大小的特定颗粒的均匀种群的条件仍然难以捉摸。然而,明确定义的均匀性以及批与批之间的高度一致性是佐剂配方用于实际疫苗的先决条件。为了完成本研究的上述任务,我们在严格控制的热力学条件下,通过动态光散射(DLS)证明了Quil-A和胆固醇形成了定义明确的蠕虫状稳定颗粒,平均直径约为40 nm, Quil-A很少或没有降解。与未配制的游离Quil-A相比,制备的纳米颗粒具有显著降低的溶血作用。然而,当同样的条件应用于QS-21/胆固醇纳米颗粒形成时,所获得的形貌是平均直径约为70 nm的螺旋弹簧状颗粒。这种颗粒形态和大小的差异归因于Quil-A和QS-21的疏水性差异,Quil-A是酰基化和去酰基化皂苷的异种组分。随着工艺优化导致标准化的颗粒大小分布和纳米颗粒形态,NanoQuil F70具有作为一种良好耐受的疫苗佐剂的潜力,可用于兽医,提高Th1/Th2平衡的免疫反应。
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