Synthesis of PEG-PLA Block Copolymer Nanospheres Containing Mixed Recombinant Proteins of Protective Antigen and Lethal Factor of Bacillus anthracis

H. Honari, M. Minaei, Hassan Mirhaj, Sayyed Masih Etemadaubi
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Abstract

Article type: Orginal article Introduction: Anthrax is a zoonotic disease and vaccine production is one way to protect people against this bacterium. This study investigates the application of copolymer nanocapsule (PEG-PLA) systems with controlled release specificity for a recombinant protective antigen and lethal factor of Bacillus anthracis to provide a vaccine candidate. Material & Methods: In this experimental study, the dual solvent evaporation emulsion method was used to produce nanocapsules. Moreover, zeta potential and size of nanoparticles, loading efficiency of nanoparticles, recombinant protein release pattern, the probable effect of PLA-PEG nanoparticle production on the shelf life of recombinant proteins were investigated in this study. Mice were used as test and control samples for antibody production and immune response evaluation. (Ethic code: 9727250) Findings: The results of this study showed that mixed proteins (LFD1+PAD4) loaded on PEG-PLA block copolymer had an average size of 109 nm, the zeta potential of -27.7 mV, and PDI=0.394. In this study, it was shown that the release of these antigens was carried out in two stages of rapid and slow release. The release of the proteins was estimated at about 20% on the first day and 78% on the 49th day. The titers of antibodies produced in the serum of the mouse groups against these antigens at defined intervals were significantly different from each other. Discussion & Conclusion: The results of this study suggest the application of PEG-PLA block copolymer nanocapsules containing mixed recombinant proteins of protective antigen and lethal factor of Bacillus anthracis. Due to less degradation and more protection of antigen activity in nanoparticles, compared to traditional methods, as well as fewer repeat injections, higher specificity, reduced side effects, lower cost, and slower release rate, the use of nanoparticles is a good option to replace traditional methods.
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炭疽芽孢杆菌保护抗原与致死因子混合重组蛋白聚乙二醇-聚乳酸嵌段共聚物纳米球的合成
文章类型:原文章简介:炭疽是一种人畜共患疾病,疫苗生产是保护人们免受这种细菌侵害的一种方法。本研究研究了共聚物纳米胶囊(PEG-PLA)系统在炭疽芽孢杆菌保护抗原和致死因子的控释特异性的应用,以提供一种候选疫苗。材料与方法:本实验采用双溶剂蒸发乳液法制备纳米胶囊。此外,本研究还考察了纳米颗粒的zeta电位和大小、纳米颗粒的负载效率、重组蛋白的释放模式以及PLA-PEG纳米颗粒的制备对重组蛋白保质期的可能影响。用小鼠作为试验和对照样品进行抗体产生和免疫应答评价。(伦理代码:9727250)结果表明:PEG-PLA嵌段共聚物上负载的混合蛋白(LFD1+PAD4)平均粒径为109 nm, zeta电位为-27.7 mV, PDI=0.394。本研究表明,这些抗原的释放分为快速释放和缓慢释放两个阶段。据估计,第一天蛋白质的释放量约为20%,第49天为78%。在确定的时间间隔内,小鼠血清中针对这些抗原产生的抗体滴度彼此显著不同。讨论与结论:本研究结果提示含有炭疽芽孢杆菌保护抗原与致死因子混合重组蛋白的PEG-PLA嵌段共聚物纳米胶囊的应用前景广阔。由于与传统方法相比,纳米颗粒降解更少,对抗原活性的保护更强,并且重复注射次数更少,特异性更高,副作用更小,成本更低,释放速度更慢,因此使用纳米颗粒是替代传统方法的一个很好的选择。
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