Luo Hong, Ma Yanyan, Su Zhiguo, Gu Yanhao, Zhang Songping, Gerstweiler Lukas
School of Chemical Engineering, Faculty of Science, Engineering and Technology, University of Adelaide, Adelaide 5005, Australia; State Key Laboratory of Biochemical Engineering, Key Laboratory of Biopharmaceutical Preparation and Delivery (CAS), Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; Institute of Pharmaceutical and Food Engineering, Shanxi University of Chinese Medicine, Jinzhong 030619, PR China.
State Key Laboratory of Biochemical Engineering, Key Laboratory of Biopharmaceutical Preparation and Delivery (CAS), Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
Int J Biol Macromol. 2025 Jan;286:138372. doi: 10.1016/j.ijbiomac.2024.138372. Epub 2024 Dec 4.
The development of modular virus-like particle (VLP) vaccine platforms with genetically inserted antigens in viral structural proteins shows great promise for advancing vaccine technology. However, the instability of many constructs leads to trial-and-error approaches, and the challenge of predicting stability based solely on amino acid sequences remains unresolved, yet highly appealing. This study evaluates the stability of wild-type murine polyomavirus (MPV) VP1 capsomeres and three engineered chimeric variants using molecular dynamics (MD) simulations and laboratory experiments. MD simulations, based on AlphaFold2 predictions and up-to-date all-atom force fields, accurately predicted the thermal stability and hydrophobicity of VP1-based capsomeres. Thermodynamic analysis revealed that binding energies from simulations reliably indicate thermal stability. Experiments and simulation results showed that inserts influence the stability of capsomeres differently, with larger insertions generally having a greater impact on the structures of capsomeres. This leads to increased intra-subunit distances and a higher proportion of flexible regions in the capsomere chassis. Capsomeres with less compact structures were found to have lower thermal stability. Specifically, the thermal transitional temperature (T) of the wild-type capsomeres was 46.9 °C, while the T values of the three chimeric derivatives were 42.0 °C, 38.8 °C, and 37.7 °C, reflecting a correlation between decreased thermal stability and reduced structural compactness. This research presents a robust approach for predicting the stability of novel VLP constructs based on amino acid sequences, potentially enhancing vaccine design by reducing failures, and suggests a shift towards minimal epitope insertions for improved stability.
在病毒结构蛋白中通过基因插入抗原开发模块化病毒样颗粒（VLP）疫苗平台，在推进疫苗技术方面显示出巨大潜力。然而，许多构建体的不稳定性导致了反复试验的方法，仅基于氨基酸序列预测稳定性的挑战仍未解决，但却极具吸引力。本研究使用分子动力学（MD）模拟和实验室实验评估了野生型小鼠多瘤病毒（MPV）VP1 衣壳粒和三种工程化嵌合变体的稳定性。基于 AlphaFold2 预测和最新全原子力场的 MD 模拟准确预测了基于 VP1 的衣壳粒的热稳定性和疏水性。热力学分析表明，模拟得到的结合能可靠地指示了热稳定性。实验和模拟结果表明，插入片段对衣壳粒稳定性的影响各不相同，较大的插入片段通常对衣壳粒结构有更大影响。这导致衣壳粒底盘内亚基间距离增加，柔性区域比例更高。发现结构不太紧凑的衣壳粒热稳定性较低。具体而言，野生型衣壳粒的热转变温度（T）为 46.9℃，而三种嵌合衍生物的 T 值分别为 42.0℃、38.8℃和 37.7℃，这反映了热稳定性降低与结构紧凑性降低之间的相关性。本研究提出了一种基于氨基酸序列预测新型 VLP 构建体稳定性的可靠方法，有望通过减少失败来加强疫苗设计，并建议转向最小化表位插入以提高稳定性。
