Athirathinam Krubha, Nandakumar Selvasudha, Verma Shailendra Kumar, Kandasamy Ruckmani
Department of Pharmaceutical Technology, Centre for Excellence in Nano-Bio Translational Research (CENTRE), University College of Engineering, Bharathidasan Institute of Technology, Anna University, Tiruchirappalli 620024, India.
Department of Biotechnology, Pondicherry University, Puducherry Union Territory, Puducherry 605014, India.
Vaccines (Basel). 2022 Dec 23;11(1):27. doi: 10.3390/vaccines11010027.
Generally, protein-based vaccines are available in liquid form and are highly susceptible to instability under elevated temperature changes including freezing conditions. There is a need to create a convenient formulation of protein/peptides that can be stored at ambient conditions without loss of activity or production of adverse effects. The efficiency of naturally occurring biocompatible polymer dextran in improving the shelf-life and biological activity of a highly thermally unstable plague vaccine candidate protein called Low Calcium Response V antigen (LcrV), which can be stored at room temperature (30 ± 2 °C), has been evaluated. To determine the preferential interactions with molecular-level insight into solvent-protein interactions, analytical techniques such asspectroscopy, particle size distribution, gel electrophoresis, microscopy, and thermal analysis have been performed along with the evaluation of humoral immune response, invivo. The analytical methods demonstrate the structural stability of the LcrV protein by expressing its interaction with the excipients in the formulation. The invivo studies elicited the biological activity of the formulated antigen with a significantly higher humoral immune response (-value = 0.047) when compared to the native, adjuvanted antigen. We propose dextran as a potential biopolymer with its co-excipient sodium chloride (NaCl) to provide protein compactness, i.e., prevent protein unfolding by molecular crowding or masking mechanism using preferential hydrophobic interaction for up to three weeks at room temperature (30 ± 2 °C).
一般来说，基于蛋白质的疫苗为液体形式，在包括冷冻条件在内的温度升高变化下极易变得不稳定。需要创建一种方便的蛋白质/肽制剂，其能够在环境条件下储存而不丧失活性或产生不良反应。已经评估了天然存在的生物相容性聚合物葡聚糖在提高一种名为低钙反应V抗原（LcrV）的高度热不稳定鼠疫疫苗候选蛋白的保质期和生物活性方面的效率，该蛋白可以在室温（30±2°C）下储存。为了通过对溶剂-蛋白质相互作用的分子水平洞察来确定优先相互作用，已进行了诸如光谱学、粒度分布、凝胶电泳、显微镜检查和热分析等分析技术，同时还评估了体内体液免疫反应。分析方法通过表达LcrV蛋白与制剂中辅料的相互作用来证明其结构稳定性。与天然佐剂化抗原相比，体内研究引发了配制抗原的生物活性，其体液免疫反应显著更高（P值=0.047）。我们提出葡聚糖与其辅料氯化钠（NaCl）作为一种潜在的生物聚合物，以提供蛋白质致密性，即通过分子拥挤或屏蔽机制，利用优先疏水相互作用在室温（30±2°C）下防止蛋白质展开长达三周。
