Shayesteh Mohadeseh, Ghasemi Fahimeh, Tabandeh Fatemeh, Yakhchali Bagher, Shakibaie Mehdi
Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, I.R. Iran.
Department of Medical Biotechnology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, I.R. Iran.
Res Pharm Sci. 2020 May 11;15(2):144-153. doi: 10.4103/1735-5362.283814. eCollection 2020 Apr.
Codon optimization has been considered as a powerful strategy to increase the expression level of protein therapeutics in mammalian cells. As an empirical approach to study the effects of the codon usage and GC content on heterologous gene expression in suspension adapted Chinese hamster ovary (CHO-s) cells, we redesigned the recombinant human interferon beta (rhIFN- β) gene based on the codon preference of the CHO cell in a way to increase the GC content in the third position of each codon.
The nucleotide sequence of the codon-optimized rhIFN-β was synthesized in parallel with the wild-type and expressed transiently in CHO-s cells using Epstein-Bar virus (EBV)-based expression system. The protein expression of the rhIFN-β by codon-optimized and wild-type genes were quantified using ELISA test.
FINDINGS / RESULTS: The results indicated a 2.8-fold increase in the expression level of the biologically active form of the rhIFN-β by codon-optimized sequence.
These results shed light on the capability of codon optimization to create a stable CHO cell for scaling up the production of recombinant therapeutics such as rhIFN-β.
密码子优化被认为是提高蛋白质治疗药物在哺乳动物细胞中表达水平的一种有效策略。作为一种研究密码子使用和GC含量对悬浮适应型中国仓鼠卵巢细胞（CHO-s）中异源基因表达影响的经验性方法，我们根据CHO细胞的密码子偏好重新设计了重组人干扰素β（rhIFN-β）基因，以增加每个密码子第三位的GC含量。
密码子优化的rhIFN-β的核苷酸序列与野生型序列并行合成，并使用基于爱泼斯坦-巴尔病毒（EBV）的表达系统在CHO-s细胞中瞬时表达。使用ELISA试验对密码子优化基因和野生型基因表达的rhIFN-β蛋白进行定量。
结果表明，密码子优化序列使rhIFN-β生物活性形式的表达水平提高了2.8倍。
这些结果揭示了密码子优化在创建稳定的CHO细胞以扩大重组治疗药物如rhIFN-β生产方面的能力。
