GSK, Siena, Italy.
GSK, Siena, Italy.
Vaccine. 2020 Apr 23;38(19):3600-3609. doi: 10.1016/j.vaccine.2020.02.001. Epub 2020 Feb 13.
Aluminum based adjuvants are widely used in commercial vaccines, since they are known to be safe and effective with a variety of antigens. The effect of antigen adsorption onto Aluminum Hydroxide is a complex area, since several mechanisms are involved simultaneously, whose impact is both antigen and formulation conditions dependent. Moreover, the mode of action of Aluminum Hydroxide is itself complex, with many mechanisms operating simultaneously. Within the literature there are contrasting theories regarding the effect of adsorption on antigen integrity and stability, with reports of antigen being stabilized by adsorption onto Aluminum Hydroxide, but also with contrary reports of antigen being destabilized. With the aim to understand the impact of adsorption on three recombinant proteins which, following in vivo immunization, are able to induce functional bactericidal antibodies against Neisseria meningitidis type B, we used a range of physico-chemical tools, such as DSC and UPLC, along with in vitro binding of antibodies that recognize structural elements of the proteins, and supported the in vitro data with in vivo evaluation in mice studies. We showed that, following exposure to accelerated degradation conditions involving heat, the recombinant proteins, although robust, were stabilized by adsorption onto Aluminum Hydroxide and retain their structural integrity unlike the not adsorbed proteins. The measure of the Melting Temperature was a useful tool to compare the behavior of proteins adsorbed and not adsorbed on Aluminum Hydroxide and to predict protein stability.
铝佐剂被广泛应用于商业疫苗中，因为它们被认为是安全有效的，可与多种抗原结合。抗原吸附到氢氧化铝上的效果是一个复杂的领域，因为有几个机制同时起作用，其影响既取决于抗原又取决于制剂条件。此外，氢氧化铝的作用方式本身也很复杂，有许多机制同时起作用。在文献中，关于吸附对抗原完整性和稳定性的影响存在相互矛盾的理论，有报道称抗原通过吸附到氢氧化铝上而稳定，但也有相反的报道称抗原不稳定。为了了解吸附对三种重组蛋白的影响，这三种重组蛋白在体内免疫接种后能够诱导针对脑膜炎奈瑟菌 B 型的功能性杀菌抗体，我们使用了一系列物理化学工具，如 DSC 和 UPLC，以及识别蛋白质结构元素的抗体的体外结合，并通过体内评估在小鼠研究中支持体外数据。我们表明，在经历涉及热的加速降解条件后，尽管重组蛋白很稳定，但通过吸附到氢氧化铝上而得到稳定，并保持其结构完整性，而未吸附的蛋白质则不然。测量熔点是一种有用的工具，可以比较吸附和未吸附在氢氧化铝上的蛋白质的行为，并预测蛋白质的稳定性。
