摘要
构建弓形虫多表位PLGA纳米疫苗并评估其可行性,预测弓形虫TgMIC3蛋白的B、T淋巴细胞优势抗原表位,联合抗原表位SAG1((238-256))、GRA7((20-28))和AS15,构建出新型多表位肽,用生物信息学工具对其理化性质、三级结构和分子对接能力等特征进行分析;原核表达并纯化多表位重组蛋白(multi-epitope protein, MEP),通过Western blot鉴定纯化效果;搭建MEP-PLGA纳米递送体系,对纳米颗粒进行表征。结果表明,多表位肽为亲水性蛋白非过敏原,具有一定抗原性,抗原表位均呈现在蛋白表面,能与MHC分子H-2-Dd稳定结合;可在BL21(DE3)大肠埃希氏菌表达系统中稳定表达,并能被特异性抗体所识别;构建的MEP-PLGA纳米递送体系包封为54.42%,纳米颗粒饱满,大小均一。成功设计并构建出弓形虫多表位PLGA纳米疫苗,为弓形虫新型纳米疫苗的研发奠定了基础。
This study aimed to design a PLGA nanoparticle with multi-epitopes against Toxoplasma gondii and evaluate its feasibility for vaccination.The dominant T and B cell epitopes of TgMIC3 protein was predicted.A novel peptide with multi-epitope was constructed with epitopes of TgMIC3,SAG1(238-256),GRA7(20-28)and AS15.Bioinformatic tools were applied to analyze the physicochemical features,tertiary structures and molecular docking of the novel peptide.The Multi-epitope protein(MEP)was obtained by prokaryotic expression and identified by Western blot.The MEP-PLGA nanoparticles delivering system was synthesized and characterized.The results suggested that MEP was a hydrophilic protein and non-allergen with certain immunogenicity.The epitopes were present on the surface of the protein,which could stably bind with MHC molecule H-2-Dd.MEP could be stably expressed in the BL21(DE3)Escherichia coli expression system and recognized by specific antibodies.The encapsulation efficiency of MEP-PLGA was measured as 54.42%with full and uniform nanoparticles in size.A MEP-PLGA nanoplatform for vaccination against T.gondii was successfully designed and constructed,laying the foundation for the development of a new type of T.gondii nanovaccine.
作者
张飞跃
伍孟琛
吴海燕
马光旭
吴飞
杜爱芳
杨怡
ZHANG Fei-yue;WU Meng-chen;WU Hai-yan;MA Guang-xu;WU Fei;DU Ai-fang;YANG Yi(Institute of Preventive Veterinary Medicine,Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine,Zhejiang University,Hangzhou,Zhejiang,310058,China)
出处
《动物医学进展》
北大核心
2024年第2期1-6,共6页
Progress In Veterinary Medicine
基金
浙江省自然科学基金项目(LQ21C180002)
中国博士后科学基金项目(2022M722788)。
关键词
弓形虫
多表位重组蛋白
PLGA纳米疫苗
Toxoplasma gondii
multi-epitope recombinant protein
PLGA nanovaccine
