Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells.

Q2 Medicine Antibody Therapeutics Pub Date : 2022-10-01 DOI:10.1093/abt/tbac025

Han Kee Ong, Ngan T B Nguyen, Jiawu Bi, Yuansheng Yang

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引用次数: 2

Abstract

Background: Two-armed FabscFv-Fc is a favoured bispecific antibody (BsAb) format due to its advantages of the conventional IgG structure. Production of FabscFv-Fc requires expression of three polypeptide chains, one light chain (LC), one heavy chain (HC) and a scFv fused to the Fc (scFvFc) at optimal ratios.

Methods: We designed a set of internal ribosome entry site (IRES)-mediated multi-cistronic vectors tailoring to various expression ratios of the three polypeptides to study how the chain ratios affect the FabscFv-Fc production.

Results: Expression of HC and scFvFc chains at 1:1 ratio and excess LC gave the highest yield of correctly assembled product. Compared to the use of IRES and multiple promoters, using 2A peptides for co-expression of the three polypeptides gave the highest titre and correctly assembled product.

Conclusion: The results obtained in this work provide insights to the impacts of hetero-chain ratios on the BsAb production.

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提高CHO细胞中基于旋钮入孔的FabscFv-Fc双特异性抗体表达水平和组装的载体设计。

背景:双臂FabscFv-Fc是一种受欢迎的双特异性抗体(BsAb)格式，由于其传统IgG结构的优点。生产FabscFv-Fc需要表达三条多肽链，一条轻链(LC)，一条重链(HC)和一个以最佳比例融合到Fc的scFv (scFvFc)。方法:我们设计了一套内部核糖体进入位点(IRES)介导的多顺反子载体，根据三种多肽的不同表达比例剪裁，研究链比例对FabscFv-Fc产生的影响。结果:HC和scFvFc链以1:1的比例表达，过量的LC使正确组装的产物产量最高。与使用IRES和多个启动子相比，使用2A肽共表达三种多肽可获得最高滴度和正确组装的产物。结论:本研究结果揭示了杂链比例对BsAb生成的影响。

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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