五株大肠杆菌在BenchtopBioreactor中获得由与MAP30毒素结合的VEGF抗体组成的重组免疫毒素的最大产量的比较

Q3 Biochemistry, Genetics and Molecular Biology
Mina Zarei, Mohammad Hossein Morowvat
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引用次数: 0

摘要

癌症是世界范围内的主要死亡原因之一,给所有社会的卫生系统带来了高昂的成本。需要广泛的生物学研究来发现合适的治疗方法。大肠杆菌一直被认为是生产重组蛋白治疗药物的主要生物技术生物工厂之一。在本研究中,研究人员比较了5株大肠杆菌,以在台式生物反应器中最大限度地生产先前设计的携带免疫毒素的重组MAP30毒素,以对抗vegf过表达的癌细胞。从encbi数据库中提取重组免疫毒素编码基因序列。用于产生重组免疫毒素的宿主为BL21 (DE3)、DH5α、SHuffle®T7、XL1-Blue和Rosetta-gamiTM (DE3) 5种大肠杆菌。用cacl2法进行细菌转化。使用600 nm光密度测量进行细菌生长测量。采用SDS-PAGE分析测定免疫毒素的产生。在10- l台式搅拌槽生物反应器中培养出产量最高的菌株。结果表明,与其他菌株相比,BL21 (DE3)菌株的重组蛋白表达量最高。此外,大肠杆菌大肠杆菌21 (DE3)和SHuffle®T7在LB培养基中转化前的细胞生长显著高于其他菌株。此外,Rosetta-gami的转化与细胞增殖减少有关。XL1-Blue菌株的转化对细胞生长没有影响。生长动力学分析表明转化后的BL21 (DE3)细胞在实验室台式生物反应器中有适当的增殖。基于本研究结果,BL21 (DE3)菌株可作为在搅拌槽生物反应器中生产抗VEGF重组免疫毒素的合适宿主,用于肿瘤的治疗。然而,它的确切机制还有待于广泛的研究。基于本研究结果,BL21 (DE3)菌株可作为搅拌式生物反应器中生产抗VEGF重组免疫毒素的合适宿主,用于肿瘤的治疗。然而,它的确切机制还有待于广泛的研究。没有一个
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Five Escherichia coli Strains to Achieve the Maximum Yield of a Recombinant Immunotoxin Consisting of an Antibody against VEGF Conjugated with MAP30 Toxin in a Benchtop Bioreactor.

Background: Cancer is among the leading causes of death worldwide, imposing high costs on the health systems of all societies. Extensive biological studies are required to discover appropriate therapies. Escherichia coli has long been regarded as one of the main biotechnological bio-factories to produce recombinant protein-based therapeutics. In the present study, five strains of E. coli were compared to achieve the maximum production of a previously designed recombinant immunotoxin-carrying MAP30 toxin against VEGF-overexpressed cancer cells in a benchtop bioreactor.

Methods: The recombinant immunotoxin coding gene sequence was extracted from the NCBI database. The host used to produce the recombinant immunotoxin were five E. coli strains of BL21 (DE3), DH5α, SHuffle®T7, XL1-Blue, and Rosetta-gamiTM (DE3). CaCl2 method was used for bacterial transformation. Bacterial growth measurements were performed using optical density measurements at 600 nm. The immunotoxin production was measured using SDS-PAGE analysis. The best-producing strain was cultivated in a 10-L benchtop stirred tank bioreactor. Recent patents on this field were also studied.

Results: The results demonstrated that the BL21 (DE3) strain had the highest expression of recombinant protein in comparison to other strains. Moreover, the cell growth of E. coli BL21 (DE3) and SHuffle®T7 strains before transformation in the LB medium, were significantly higher in comparison to other strains. Additionally, the transformation of Rosettagami was associated with decreased cell proliferation. The transformation of the XL1-Blue strain did not effect cell growth. Analysis of the growth kinetics demonstrated appropriate proliferation of the transformed BL21 (DE3) cells in the laboratory benchtop bioreactor.

Conclusions: Based on the results of this study, the BL21 (DE3) strain could be used as a suitable host for the production of the recombinant immunotoxin against VEGF in stirred tank bioreactor, which can be employed for the treatment of tumors. Yet, its precise mechanism must be explored in extensive studies.

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来源期刊
Recent patents on biotechnology
Recent patents on biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
2.90
自引率
0.00%
发文量
51
期刊介绍: Recent Patents on Biotechnology publishes review articles by experts on recent patents on biotechnology. A selection of important and recent patents on biotechnology is also included in the journal. The journal is essential reading for all researchers involved in all fields of biotechnology.
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