A Comparative Investigation of the Bispecific Antibody: Expression in Expi293F Cells and E.coli

R. Moazzami, Hasan Mirzahosaini, F. Naddafi, Fatemeh Davami
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Abstract

Aims: Dual-Character antibodies can simultaneously target two surface markers. Blinatumomab is a C19 / CD3 antibody from the BiTE family (Bispecific T cell engager antibody) and was approved by the US Food and Drug Administration for clinical use. This antibody effectively targets malignant cells in patients with acute infoblastic leukemia. In the production of large quantities of such antibodies on an industrial scale, selecting the appropriate host remains a critical issue. Mammalian cells and strains of E.coli are the most common hosts for producing antibodies and antibody components on an industrial scale, respectively. Methods & Materials: In this study, a dual-specific antibody was used in the mammalian system of plasmid pcDNA3.1 (+) and for expression in the bacterial system of plasmid pET22b. The antibody produced in both systems was purified using nickel affinity resin under similar conditions. Next, SDS-PAGE and Western blot analysis was performed on both study samples. Finally, the binding properties of the antibody secreted from both systems were assessed by the ELISA test. Findings: The present study results suggested that antibodies produced by the mammalian expression system provided better binding properties than the expression system in bacteria. Conclusion: This study indicated that in the case of antibodies to two traits of the BiTE family, like Blinatombe, mammalian cells generate a more efficient and successful expression system; although the bacterium can produce much larger amounts of the antibody.
双特异性抗体在Expi293F细胞和大肠杆菌中表达的比较研究
目的:双特性抗体可同时靶向两种表面标记物。Blinatumomab是一种来自BiTE家族(双特异性T细胞接触抗体)的C19 / CD3抗体,已被美国食品和药物管理局批准临床使用。该抗体有效靶向急性非母细胞白血病患者的恶性细胞。在工业规模上大量生产这种抗体时,选择合适的宿主仍然是一个关键问题。哺乳动物细胞和大肠杆菌菌株分别是在工业规模上生产抗体和抗体成分的最常见宿主。方法与材料:采用双特异性抗体在哺乳动物质粒pcDNA3.1(+)和细菌质粒pET22b中表达。在相似的条件下,用镍亲和树脂纯化两种体系产生的抗体。接下来,对两个研究样本进行SDS-PAGE和Western blot分析。最后,通过ELISA测试评估两种系统分泌的抗体的结合特性。结果:本研究结果表明,哺乳动物表达系统产生的抗体比细菌表达系统具有更好的结合特性。结论:本研究表明,在针对BiTE家族两种特征(如Blinatombe)的抗体的情况下,哺乳动物细胞产生了更有效和成功的表达系统;虽然细菌可以产生大量的抗体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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