Preparation and Evaluation of Rivax Protein Loading in Chitosan Nanoparticles

Davood Sadeghi, F. Ebrahimi, M. Zeinoddini, Y. Tarverdizadeh, Mostafa Bakhshi, MJ Bageripor, SM Agaii
{"title":"Preparation and Evaluation of Rivax Protein Loading in Chitosan Nanoparticles","authors":"Davood Sadeghi, F. Ebrahimi, M. Zeinoddini, Y. Tarverdizadeh, Mostafa Bakhshi, MJ Bageripor, SM Agaii","doi":"10.18869/ACADPUB.JBUMS.20.6.62","DOIUrl":null,"url":null,"abstract":"BACKGROUND AND OBJECTIVE: Ricin toxin is a heterodimer glycoprotein which, due to its high toxicity, is used as a bioterrorism agent. Immunogenicity studies against ricin are now focused on two subunit vaccine candidates, including RiVax and RVEc. These studies have examined the vaccine candidate immunization as an alone and in combination with adjuvant, however, there is not a published study on the immunogenicity evaluation of the candidate vaccine through the delivery by nanoparticles. The aim of this study was preparation and evaluation of RiVax recombinant vaccine-loading in chitosan nanoparticles. METHODS: In this experimental study, After transferring the RiVax gene to the bacterium, inducing the expression and purification of the RiVax protein by affinity chromatography column, the RiVax protein was loaded with Ionic Gelation method in chitosan nanoparticles. Then, the properties of nanoparticles including size, morphology, loading percentage and release pattern of RiVax protein from nanoparticles and stability of this protein during acidic loading conditions in nanoparticles by SDS-PAGE were evaluated. Also, Immunization study were performed on 3 mouse groups (n=4/group) by RiVax protein, Nanoparticles containing protein and phosphate buffer. FINDING: The results of this study showed that the nanoparticles containing protein had a size of 178 nm and a Zeta potential of +27.8 MV and a polydispersity index of 0.193. Also, according to SDS-PAGE results, it was found that the RiVax recombinant protein was denatured during the process of preparing the chitosan nanoparticles. CONCLUSION: The results of this study showed that the RiVax protein has been unstable in acidic conditions for the production of chitosan nanoparticles and Ionic Gelation method is not suitable for loading this protein in chitosan nanoparticles.","PeriodicalId":15108,"journal":{"name":"Journal of Babol University of Medical Sciences","volume":"48 1","pages":"62-69"},"PeriodicalIF":0.0000,"publicationDate":"2018-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Babol University of Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.18869/ACADPUB.JBUMS.20.6.62","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0

Abstract

BACKGROUND AND OBJECTIVE: Ricin toxin is a heterodimer glycoprotein which, due to its high toxicity, is used as a bioterrorism agent. Immunogenicity studies against ricin are now focused on two subunit vaccine candidates, including RiVax and RVEc. These studies have examined the vaccine candidate immunization as an alone and in combination with adjuvant, however, there is not a published study on the immunogenicity evaluation of the candidate vaccine through the delivery by nanoparticles. The aim of this study was preparation and evaluation of RiVax recombinant vaccine-loading in chitosan nanoparticles. METHODS: In this experimental study, After transferring the RiVax gene to the bacterium, inducing the expression and purification of the RiVax protein by affinity chromatography column, the RiVax protein was loaded with Ionic Gelation method in chitosan nanoparticles. Then, the properties of nanoparticles including size, morphology, loading percentage and release pattern of RiVax protein from nanoparticles and stability of this protein during acidic loading conditions in nanoparticles by SDS-PAGE were evaluated. Also, Immunization study were performed on 3 mouse groups (n=4/group) by RiVax protein, Nanoparticles containing protein and phosphate buffer. FINDING: The results of this study showed that the nanoparticles containing protein had a size of 178 nm and a Zeta potential of +27.8 MV and a polydispersity index of 0.193. Also, according to SDS-PAGE results, it was found that the RiVax recombinant protein was denatured during the process of preparing the chitosan nanoparticles. CONCLUSION: The results of this study showed that the RiVax protein has been unstable in acidic conditions for the production of chitosan nanoparticles and Ionic Gelation method is not suitable for loading this protein in chitosan nanoparticles.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
壳聚糖纳米颗粒负载Rivax蛋白的制备及性能评价
背景与目的:蓖麻毒素是一种异源二聚体糖蛋白,由于其高毒性,被用作生物恐怖制剂。针对蓖麻毒素的免疫原性研究目前集中在两种亚单位候选疫苗上,包括RiVax和RVEc。这些研究检查了候选疫苗单独免疫和与佐剂联合免疫,然而,没有关于通过纳米颗粒递送的候选疫苗免疫原性评价的发表研究。本研究的目的是制备和评价壳聚糖纳米颗粒负载RiVax重组疫苗。方法:在本实验研究中,将RiVax基因转移到细菌体内,通过亲和层析柱诱导RiVax蛋白的表达和纯化,用离子凝胶法将RiVax蛋白装入壳聚糖纳米颗粒中。然后,利用SDS-PAGE评价了RiVax蛋白在纳米颗粒酸性负载条件下的稳定性,包括纳米颗粒的大小、形态、负载率和释放模式。采用RiVax蛋白、含蛋白纳米颗粒和磷酸盐缓冲液对3组小鼠(n=4/组)进行免疫实验。发现:本研究结果表明,含蛋白质的纳米颗粒尺寸为178 nm, Zeta电位为+27.8 MV,多分散性指数为0.193。此外,SDS-PAGE结果表明,在制备壳聚糖纳米颗粒的过程中,RiVax重组蛋白发生变性。结论:本研究结果表明,RiVax蛋白在酸性条件下制备壳聚糖纳米颗粒是不稳定的,离子凝胶法不适合在壳聚糖纳米颗粒中负载该蛋白。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
CiteScore
0.50
自引率
0.00%
发文量
1
审稿时长
12 weeks
期刊最新文献
نقش پیشگویی کننده لکوسیتوز، آنمی و ترومبوسیتوز قبل از جراحی با شدت تومورهای اپیتلیالی تخمدان ژن های عامل بیماری ادرار شربت افرا ، پاتو مکانیسم مولکولی بیماری و جهش های جمعیت ایران Nurses’ Experience of Pain Management in Patients Referred to the Emergency Department A Case Report of Mucopolysaccharidosis Type VI Maternal Exposure to Air Pollution and Fetal Abnormalities
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1