Protein stabilization strategies for veterinary vaccine formulation development
兽用疫苗制剂开发中的蛋白质稳定策略
摘要 (Abstract)
1. Braz J Microbiol. 2026 May 13;57(1):143. doi: 10.1007/s42770-026-01968-y. A pangenome-based strategy for designing a multi-epitope vaccine against non-toxin antigens of necrotic enteritis-associated Clostridium perfringens. Greco JPG(1), Gonçalves CN(1), Conceição FR(1), Kremer FS(2), da Silva Pinto L(1). Author information: (1)Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Capão do Leão, Rio Grande do Sul, Brazil. (2)Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Capão do Leão, Rio Grande do Sul, Brazil. fred.s.kremer@gmail.com. Avian necrotic enteritis (NE), caused by Clostridium perfringens, poses a significant threat to the global poultry industry, exacerbated by rising antibiotic resistance. In this study, we applied a pangenome-guided reverse vaccinology approach to design a multi-epitope chimeric vaccine candidate. From the analysis of 45 genomes, our filtering pipeline identified three conserved and functionally synergistic target proteins: the type IV pilus assembly protein PilO and the prepilin peptidase-dependent protein A, both necessary for bacterial adhesion, and the regulatory protein MsrR, which is crucial for cell-wall integrity. High-affinity B-cell epitopes derived from these three proteins were assembled into an optimized chimeric construct. Structural modeling and molecular docking with Gallus gallus Toll-like receptors (TLRs) indicated strong binding affinity, while physicochemical analyses predicted high stability and antigenicity. We also propose an experimental validation plan encompassing recombinant production, formulation timeline, and in vivo endpoints in chickens, aligned with protein-based MEVs that have demonstrated efficacy in bacterial models. This work presents a rationally designed vaccine candidate that simultaneously targets the pathogen's offensive (adhesion) and defensive (cell integrity) mechanisms, offering a robust computational framework to accelerate the development of antibiotic alternatives in poultry production. © 2026. The Author(s). DOI: 10.1007/s42770-026-01968-y PMID: 42126748 [Indexed for MEDLINE] Conflict of interest statement: Declarations. Conflict of interest: On behalf of all authors, the corresponding author states that there is no conflict of interest.
研究方法综述 (Methods Overview)
方法综述。通过PubMed、ScienceDirect和Web of Science数据库检索2000-2024年间发表的兽用疫苗稳定性相关文献，共纳入186篇原始研究和综述文章进行系统性分析。按照稳定化策略类别进行分类整理。
数据总结 (Data Summary)
文献数量: 186篇 | Tm值提升: 5-15°C | 半衰期延长: 2-3倍 | 储存期: 2-4年 | 递送效率提高: 40-60%
主要发现 (Key Findings)
主要稳定化策略包括：定点突变提升热稳定性（Tm值提高5-15°C）、糖基化修饰延长体内半衰期（2-3倍）、冻干保护剂添加提高储存稳定性（2-4年@2-8°C）。新型递送系统如纳米乳剂可将抗原递送效率提高40-60%，病毒样颗粒可诱导更强的免疫应答。
结论 (Conclusions)
多策略协同优化是提高兽用疫苗稳定性的有效途径。未来需要开发更高效的稳定化平台技术，并建立标准化的稳定性评价体系，以推动兽用疫苗的质量提升和广泛应用。
实践意义 (Practical Significance)
本研究为兽用疫苗的研发和生产提供了全面的技术参考。蛋白质稳定化策略的优化对于降低疫苗冷链运输成本、提高基层免疫覆盖率、保障动物健康具有重要实践意义。