Bayat Hadi, Hossienzadeh Saghar, Pourmaleki Eśhagh, Ahani Roshanak, Rahimpour Azam
a Medical Nano-Technology and Tissue Engineering Research Center , Shahid Beheshti University of Medical Sciences , Tehran , Iran.
b Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine , Shahid Beheshti University of Medical Sciences , Tehran , Iran.
Prep Biochem Biotechnol. 2018 Feb 7;48(2):160-164. doi: 10.1080/10826068.2017.1421966. Epub 2018 Feb 9.
Monoclonal antibodies (mAbs) have emerged as the most promising category of recombinant proteins due to their high efficiency for the treatment of a wide range of human diseases. The complex nature of mAbs creates a great deal of challenges in both upstream and downstream manufacturing processes. Proportional expression and correct folding and assembly of the light chain and heavy chain are required for efficient production of the mAbs. In this regard, expression vector design has proven to have profound effects on the antibody expression level as well as its stability and quality. Here, we have explored the efficiency of different vector design strategies for the expression of a recombinant IgG1 antibody in Chinese hamster ovary (CHO) cells. The antibody expression level was analyzed in transient expression and stable cell pools followed by expression analysis on single-cell clones. While detectable amounts of antibody were observed in all three systems, dual-promoter single-vector system showed the highest expression level in transient and stable expression as well as the highest productivity among clonal cells. Our results here show the importance of vector design for successful production of whole mAbs in CHO cells.
单克隆抗体（mAbs）因其在治疗多种人类疾病方面的高效性，已成为最具前景的一类重组蛋白。单克隆抗体的复杂特性在上游和下游制造过程中都带来了诸多挑战。为了高效生产单克隆抗体，轻链和重链需要比例表达以及正确折叠和组装。在这方面，表达载体设计已被证明对抗体表达水平及其稳定性和质量有深远影响。在此，我们探讨了不同载体设计策略在中国仓鼠卵巢（CHO）细胞中表达重组IgG1抗体的效率。在瞬时表达和稳定细胞库中分析了抗体表达水平，随后对单细胞克隆进行了表达分析。虽然在所有三个系统中均观察到可检测量的抗体，但双启动子单载体系统在瞬时和稳定表达中显示出最高的表达水平，并且在克隆细胞中具有最高的生产力。我们在此的结果表明载体设计对于在CHO细胞中成功生产完整单克隆抗体至关重要。
