Malburet Camille, Leclercq Laurent, Cotte Jean-François, Thiebaud Jérôme, Cottet Hervé
IBMM, University of Montpellier, CNRS, ENSCM, Montpellier, France.
Analytical Sciences, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, Marcy-l'Étoile 69280, France.
Biomacromolecules. 2020 Aug 10;21(8):3364-3373. doi: 10.1021/acs.biomac.0c00782. Epub 2020 Jul 16.
Vaccine adjuvants are used to enhance the immune response induced by antigens that have insufficient immunostimulatory capabilities. The present work aims at developing a frontal analysis continuous capillary electrophoresis (FACCE) methodology for the study of antigen-adjuvant interactions in vaccine products. After method optimization using three cationic model proteins, namely lysozyme, cytochrome , and ribonuclease A, FACCE was successfully implemented to quantify the free antigen and thus to determine the interaction parameters (stoichiometry and binding constant) between an anionic polymeric adjuvant (polyacrylic acid, SPA09) and a cationic vaccine antigen in development for the treatment for . The influence of the ionic strength of the medium on the interactions was investigated. A strong dependence of the binding parameters with the ionic strength was observed. The concentration of the polymeric adjuvant was also found to significantly modify the ionic strength of the formulation, the extent of which could be estimated and corrected.
疫苗佐剂用于增强由免疫刺激能力不足的抗原所诱导的免疫反应。目前的工作旨在开发一种前沿分析连续毛细管电泳（FACCE）方法，用于研究疫苗产品中抗原 - 佐剂的相互作用。在用三种阳离子模型蛋白（即溶菌酶、细胞色素和核糖核酸酶A）进行方法优化后，FACCE成功用于定量游离抗原，从而确定一种阴离子聚合物佐剂（聚丙烯酸，SPA09）与一种正在开发用于治疗的阳离子疫苗抗原之间的相互作用参数（化学计量比和结合常数）。研究了介质离子强度对相互作用的影响。观察到结合参数对离子强度有很强的依赖性。还发现聚合物佐剂的浓度会显著改变制剂的离子强度，其改变程度可以估计并校正。
