枯草芽孢杆菌孢子表面展示系统保护重组蛋白免受降解——验证假说

Tomasz AAAAga, Paulina Weiher, M. Paszkiewicz, D. Nidzworski
{"title":"枯草芽孢杆菌孢子表面展示系统保护重组蛋白免受降解——验证假说","authors":"Tomasz AAAAga, Paulina Weiher, M. Paszkiewicz, D. Nidzworski","doi":"10.4172/2155-952X.1000263","DOIUrl":null,"url":null,"abstract":"Endospores of Bacillus subtilis have been used extensively as a platform for recombinant protein display for nearly two decades. Main use of the spore surface display system is generation of oral vaccines against many human and animal pathogens. Formulation of oral vaccine based on spores is an attractive approach to alternative vaccination due to the timesaving and relative easiness of production. Another advantage of such formulation is stability of presented antigens. It is assumed that the spore coat structure prevents degradation of displayed proteins by many destructive factors such as heat, proteases or stomach environment. However, there is little scientific background and substantial lack of experiments verifying this statement. In our work, we tested protective properties of spores against degradation of displayed antigens in harsh environment. We constructed B. subtilis strains producing spores presenting highly conserved long α-helix (LAH) region of the influenza A virus hemagglutinin. The constructs were obtained by fusion of LAH antigen to protein CotB or CotZ from the spore coat. We treated recombinant spores with destructive agents such as heat, protease, low pH and high-energy irradiation to test protective features of the spore coat. After treatment, spore coat extracts were analyzed by western blot to study fate of the displayed antigen. Results that we publish indicate that spore coat protects displayed antigen from degradation. This work is a strong support of hypothesis stating protective properties of spore surface display system against antigen degradation.","PeriodicalId":15156,"journal":{"name":"Journal of biotechnology & biomaterials","volume":"1 1","pages":"1-3"},"PeriodicalIF":0.0000,"publicationDate":"2017-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Bacillus subtilis Spore Surface Display System Protects Recombinant Proteins from Degradation-Verified Hypothesis\",\"authors\":\"Tomasz AAAAga, Paulina Weiher, M. Paszkiewicz, D. Nidzworski\",\"doi\":\"10.4172/2155-952X.1000263\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Endospores of Bacillus subtilis have been used extensively as a platform for recombinant protein display for nearly two decades. Main use of the spore surface display system is generation of oral vaccines against many human and animal pathogens. Formulation of oral vaccine based on spores is an attractive approach to alternative vaccination due to the timesaving and relative easiness of production. Another advantage of such formulation is stability of presented antigens. It is assumed that the spore coat structure prevents degradation of displayed proteins by many destructive factors such as heat, proteases or stomach environment. However, there is little scientific background and substantial lack of experiments verifying this statement. In our work, we tested protective properties of spores against degradation of displayed antigens in harsh environment. We constructed B. subtilis strains producing spores presenting highly conserved long α-helix (LAH) region of the influenza A virus hemagglutinin. The constructs were obtained by fusion of LAH antigen to protein CotB or CotZ from the spore coat. We treated recombinant spores with destructive agents such as heat, protease, low pH and high-energy irradiation to test protective features of the spore coat. After treatment, spore coat extracts were analyzed by western blot to study fate of the displayed antigen. Results that we publish indicate that spore coat protects displayed antigen from degradation. This work is a strong support of hypothesis stating protective properties of spore surface display system against antigen degradation.\",\"PeriodicalId\":15156,\"journal\":{\"name\":\"Journal of biotechnology & biomaterials\",\"volume\":\"1 1\",\"pages\":\"1-3\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2017-06-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of biotechnology & biomaterials\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.4172/2155-952X.1000263\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of biotechnology & biomaterials","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.4172/2155-952X.1000263","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

近二十年来,枯草芽孢杆菌的内生孢子作为重组蛋白的展示平台被广泛应用。孢子表面显示系统的主要用途是生产针对许多人类和动物病原体的口服疫苗。基于孢子的口服疫苗的配方是一种有吸引力的替代疫苗接种方法,因为节省时间和相对容易生产。这种制剂的另一个优点是所提抗原的稳定性。据推测,孢子外壳结构可以防止许多破坏性因素(如热、蛋白酶或胃环境)降解所显示的蛋白质。然而,几乎没有科学背景和大量缺乏实验来验证这一说法。在我们的工作中,我们测试了孢子在恶劣环境中对显示抗原降解的保护性能。我们构建了枯草芽孢杆菌菌株,产生具有甲型流感病毒血凝素高度保守的长α-螺旋(LAH)区域的孢子。通过将LAH抗原与孢子外壳上的CotB或CotZ蛋白融合获得了这些结构体。我们用热、蛋白酶、低pH和高能辐照等破坏剂处理重组孢子,以测试孢子外壳的保护特性。处理后,用western blot分析孢子皮提取物,研究显示抗原的命运。我们发表的结果表明,孢子外壳保护显示抗原免受降解。本研究有力地支持了孢子表面展示系统对抗原降解具有保护作用的假说。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Bacillus subtilis Spore Surface Display System Protects Recombinant Proteins from Degradation-Verified Hypothesis
Endospores of Bacillus subtilis have been used extensively as a platform for recombinant protein display for nearly two decades. Main use of the spore surface display system is generation of oral vaccines against many human and animal pathogens. Formulation of oral vaccine based on spores is an attractive approach to alternative vaccination due to the timesaving and relative easiness of production. Another advantage of such formulation is stability of presented antigens. It is assumed that the spore coat structure prevents degradation of displayed proteins by many destructive factors such as heat, proteases or stomach environment. However, there is little scientific background and substantial lack of experiments verifying this statement. In our work, we tested protective properties of spores against degradation of displayed antigens in harsh environment. We constructed B. subtilis strains producing spores presenting highly conserved long α-helix (LAH) region of the influenza A virus hemagglutinin. The constructs were obtained by fusion of LAH antigen to protein CotB or CotZ from the spore coat. We treated recombinant spores with destructive agents such as heat, protease, low pH and high-energy irradiation to test protective features of the spore coat. After treatment, spore coat extracts were analyzed by western blot to study fate of the displayed antigen. Results that we publish indicate that spore coat protects displayed antigen from degradation. This work is a strong support of hypothesis stating protective properties of spore surface display system against antigen degradation.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信