Qin-Dong Su, Min-Zhuo Guo, Yao Yi, Si-Yong Chen, Zhi-Yuan Jia, Xue-Xin Lu, Feng Qiu, Sheng-Li Bi
{"title":"[呈现preS1中和表位的两个嵌合乙肝核心颗粒的抗原分析]。","authors":"Qin-Dong Su, Min-Zhuo Guo, Yao Yi, Si-Yong Chen, Zhi-Yuan Jia, Xue-Xin Lu, Feng Qiu, Sheng-Li Bi","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.</p><p><strong>Methods: </strong>PreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.</p><p><strong>Results: </strong>The morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.</p><p><strong>Conclusion: </strong>Chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.</p>","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"336-9"},"PeriodicalIF":0.0000,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Antigenic analysis of two chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes].\",\"authors\":\"Qin-Dong Su, Min-Zhuo Guo, Yao Yi, Si-Yong Chen, Zhi-Yuan Jia, Xue-Xin Lu, Feng Qiu, Sheng-Li Bi\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.</p><p><strong>Methods: </strong>PreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.</p><p><strong>Results: </strong>The morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.</p><p><strong>Conclusion: </strong>Chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.</p>\",\"PeriodicalId\":70973,\"journal\":{\"name\":\"中华实验和临床病毒学杂志\",\"volume\":\"27 5\",\"pages\":\"336-9\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2013-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华实验和临床病毒学杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华实验和临床病毒学杂志","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的:构建含有preS1 aa 21-47表位的全长型乙型肝炎核心颗粒和表面含有preS1 aa 37-45表位的切短型乙型肝炎核心颗粒,并比较其抗原性。方法:将PreS1 aa21-47表位和aa 37-45表位分别插入全长乙肝核心(aa 1-183)和截断的HBcAg (aa 1-144),位于第78位(Asp)和第79位(Pro)之间。密码子优化后合成的基因被连接到pET43上。具有相同内聚末端(NdeI和XhoI)的1a载体,在大肠杆菌表达系统中表达。电镜观察目标蛋白的形态,ELISA和Western Blotting对其进行表征。结果:电镜下证实了病毒样颗粒的形态。H2为固体颗粒,直径为(31.61 +/- 1.27)nm; H3为空心颗粒,直径为(28.46 +/- 1.16)nm。经统计学分析,H2的直径大于H3 (P < 0.01)。采用酶联免疫吸附法(ELISA)和免疫印迹法(Western Blotting)鉴定插入抗原表位和载体蛋白的抗原性。结论:已成功表达、纯化并鉴定出表面具有preS1中和表位的乙型肝炎嵌合核心颗粒,为其在疫苗研究中的应用奠定了基础。
[Antigenic analysis of two chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes].
Objective: To construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.
Methods: PreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.
Results: The morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.
Conclusion: Chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.