Antibody titers of PEG-PLA block copolymer nanosphere containing chimeric recombinant protein of protective antigen and lethal factor of Bacillus anthracis

H. Honari, M. Minaei, H. Mirhaj, Seyyed Masih Etemad- Ayoubi
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Abstract

chimeric containing domain 1 lethal factor domain 4 protective of Bacillus anthracis in copolymer nanocapsules to the problems caused by existing vaccines and to the efficiency of the proposed Materials and Methods : In this experimental study, dual solvent evaporation method was used to produce nanocapsules containing chimeric recombinant protein of protective antigen and lethal factor of Bacillus anthracis. Zeta potential of nanoparticles, nanoparticle loading efficiency, recombinant protein release pattern, potential effect of poly (lactic acid)- poly (ethylene glycol) nanoparticle (PLA-PEG) production on the viability of recombinant proteins were investigated. Mice were used as test and control samples for antibody production and immune response evaluation. Results: The mean antibody titer produced against chimeric proteins loading was significantly different from that of free antigens. Correspondingly, the difference in antibody titer was significant between the groups of one and two times’ injection of loading and free antigens and the most antibody titer was related to two times injections of loading antigens. In addition, there was a significant difference between one times injection of loading and the free antigens. This suggests that the loading chimeric antigen on PLA-PEG nanoparticles and one time injection (instead of four times injections) could produce more antibodies. Conclusion: The results of this study showed that the domain 4 protective antigens and the domain 1 lethal factor of Bacillus anthracis could chimeric with each other and produced the active antigen. This chimeric antigen (LFD1-PA4) is active and able to inducing the animal's immune system. In addition, nanocarriers containing controlled release antigens can induce the immune system of the animal.
含炭疽芽孢杆菌保护抗原与致死因子嵌合重组蛋白的聚乙二醇-聚乳酸嵌段共聚物纳米球的抗体滴度
针对现有疫苗存在的问题以及所提出的材料和方法的有效性,本实验研究采用双溶剂蒸发法制备了含有炭疽杆菌保护抗原和致死因子嵌合重组蛋白的纳米胶囊。研究了纳米颗粒的Zeta电位、纳米颗粒的负载效率、重组蛋白的释放模式、聚乳酸-聚乙二醇纳米颗粒(PLA-PEG)的制备对重组蛋白活性的潜在影响。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。结果:嵌合蛋白的平均抗体效价与游离抗原有显著差异。相应的,1次和2次注射负载抗原和游离抗原组的抗体效价差异显著,且抗体效价最多的与2次注射负载抗原有关。此外,一次注射负载与游离抗原之间存在显著差异。这表明在PLA-PEG纳米颗粒上装载嵌合抗原并一次注射(而不是四次注射)可以产生更多的抗体。结论:炭疽芽孢杆菌的结构域4保护抗原与结构域1致死因子可以嵌合产生活性抗原。这种嵌合抗原(LFD1-PA4)具有活性,能够诱导动物的免疫系统。此外,含有控释抗原的纳米载体可以诱导动物的免疫系统。
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