合成含 BLF1 的三甲基壳聚糖纳米颗粒,并通过口服和皮下注射评估其在叙利亚小鼠中的免疫原性和保护作用。

IF 1.4 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-03-30 DOI:10.1016/j.pep.2024.106462
Ayoub fazeli, Hosein Honari, Davoud Sadeghi, Hamid Bakhtiari
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引用次数: 0

摘要

假丝酵母伯克霍尔德氏菌(Burkholderia pseudomallei)是美拉菌病(melioidosis)的病原体。在这种细菌中,BLF1 蛋白宽抑制人体细胞中蛋白质的合成。据报道,在世界某些地区,这种疾病的致死率高达 40%。目前,还没有针对这种细菌感染的有效疫苗。因此,本研究以含有 BLF1 重组蛋白的三甲基壳聚糖(TMC)聚合物为基础,合成了一种纳米疫苗,并通过口服和皮下注射对其在叙利亚小鼠中的免疫原性和保护作用进行了评估。在大肠杆菌 Bl21 (DE3) 中诱导表达 BLF1 重组蛋白,并通过亲和层析技术进行纯化。然后用离子凝胶法合成了含重组蛋白的纳米颗粒(NPs)。口服和皮下注射后,通过间接 ELISA 法评估抗体滴度。最后,使用 BLF1 毒素对小鼠组进行挑战。结果表明,皮下注射的免疫系统比口服的表现出更高的抗体滴度。然而,挑战结果却相反,口服注射的存活率更高。
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Synthesis of BLF1-containing trimethyl chitosan nanoparticles and evaluation of its immunogenicity and protection in syrian mice by oral and subcutaneous injections

The bacterium Burkholderia pseudomallei is the cause of melioidosis infectious disease. In this bacterium, the BLF1 protein wide inhibits the synthesis of proteins in human cells. This disease is reported to cause a death rate of 40% in some parts of the world. Currently, no effective vaccine is available against this bacterial infection. In this study, therefore, a Nano vaccine was synthesized based on the trimethyl chitosan (TMC) polymer containing the BLF1 recombinant protein, and its immunogenicity and protection in Syrian mice were evaluated by oral and subcutaneous injections. The BLF1 recombinant protein expression was induced in Escherichia coli Bl21 (DE3) and purified by the affinity chromatography technique. Recombinant protein-containing nanoparticles (NPs) were then synthesized by the ionotropic gelation method. After oral and subcutaneous injections, antibody titration was assessed by the indirect ELISA assay. Finally, murine groups were challenged using the BLF1 toxin. The results indicated that the immune system showed more antibody titration in subcutaneous injection than in the oral form. However, the results were reversed in the challenge results, and the survival rate was more significant in the oral injection.

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来源期刊
Protein expression and purification
Protein expression and purification 生物-生化研究方法
CiteScore
3.70
自引率
6.20%
发文量
120
审稿时长
32 days
期刊介绍: Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.
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