基于抗病毒靶向的猪繁殖与呼吸综合征病毒（PRRSV）亲和肽研究

Porcine reproductive and respiratory syndrome（PRRS）causes serious harm to the development of global pig industry, resulting in huge economic losses. At present, there are no specific medicine for PRRS, we mainly adopt a prevention-based comprehensive prevention and control measures in China, which has urgent need to develop new type antiviral agents and medicine with high performance. .Regarding PRRSV affinity peptide as the research objects in the study, PRRSV positive phage affinity were identified through the establishment of complete PRRSV particles as the target of phage peptide library screening technology, and combined with bioinformatics analysis methods to design affinity peptide sequence. Synthesized candidate PRRSV affinity peptides using chemical synthesis, PRRSV affinity peptides were identified through determination of affinity and specificity of candidate peptides and PRRSV. Affinity peptides combining with target proteins were identified through peptides-proteins interaction, and determinate the affinity of target proteins and peptides. The targeted antiviral PRRSV affinity peptides were identified through affinity peptide blocking test and their antiviral function were evaluated. The project aims to obtain PRRSV specific affinity peptides with antiviral function, lays the foundation for the research and development of new antiviral agents and medicine, and provide technical support for prevention and control of PRRS, which has significant social and economic significance and application prospects. At the same time, it also lays the theory foundation for further revealing the molecular mechanism of anti-PRRSV infection.

猪繁殖与呼吸综合征严重危害世界各国养猪业发展，造成巨大的经济损失。PRRS目前没有特效药物治疗，我国主要采取以预防为主的综合性防控措施，急需研发高效的新型抗病毒制剂和药物。本项目以PRRSV亲和肽为研究对象，通过建立以完整的PRRSV病毒粒子为靶标的噬菌体肽库筛选技术，鉴定PRRSV亲和阳性噬菌体，并结合生物信息学分析设计亲和肽序列。采用化学合成技术合成候选PRRSV亲和肽，通过测定候选多肽与PRRSV结合的亲和性和特异性，鉴定PRRSV亲和肽。利用多肽-蛋白相互作用鉴定亲和肽结合的靶蛋白并测定二者亲和力，通过亲和肽阻断试验鉴定具抗病毒靶向的PRRSV亲和肽并评价其抗病毒功能。本项目旨在获得具有抗病毒功能的PRRSV特异性亲和肽，为研发新型抗病毒制剂和药物奠定基础，为PRRS防控提供技术支撑，具有重大的社会经济意义和应用前景。同时该研究也为进一步揭示抗PRRSV感染的分子机制奠定理论基础。

猪繁殖与呼吸综合征严重危害世界各国养猪业发展，造成巨大的经济损失。PRRS目前没有特效药物治疗，我国主要采取以预防为主的综合性防控措施，急需研发高效的新型抗病毒制剂和药物。本项目以PRRSV亲和肽为研究对象，旨在研发具有抗病毒功能的PRRSV特异性亲和肽。项目实施顺利，总体进展良好，完成了各项任务并取得了一系列重要成果。通过温和纯化的新方法成功制备了结构完整、纯度高、效价高、抗原活性好、特异性强的筛选靶标PRRSV；建立PRRSV亲和肽筛选鉴定体系，获得5株PRRSV亲和阳性噬菌体；通过病毒-多肽-ELISA、间接免疫荧光、免疫过氧化物酶单层细胞检测方法，获得具有良好亲和性和特异性的PRRSV亲和肽P4；经鉴定P4结合靶蛋白为GP5，二者具有较好的亲和力；亲和肽阻断病毒感染试验证实P4为抗病毒靶向亲和肽，具有抗病毒功能。相关研究成果在国内外学术期刊上发表论文4篇，其中SCI收录3篇、中文核心1篇；获河南省科技进步二等奖1项、技术发明二等奖1项、猪蓝耳病研究进步奖1项；获授权发明专利2项，新申请发明专利6项；入选“中原英才计划—中原青年拔尖人才”和“河南省学术技术带头人”各1人，晋升正高/副高级职称共4人，培养毕业研究生3名，其中博士1名、硕士2名；项目组成员参加国内外学术会议9个共13人次，其中国际会议4个学术交流5人次，国内会议5个出席参加8人次。.该研究获得具有良好亲和性和特异性的PRRSV抗病毒靶向亲和肽P4，证实其具有抗病毒功能，且结合靶蛋白为GP5，为研发新型抗病毒制剂和药物奠定了基础，为PRRS防控提供了技术支撑，具有重大的社会经济意义和应用前景。