基于IRES序列的抗体定点整合载体构建和表达

目的：比较不同全功能抗体基因之间的表达量。方法：构建了基于FLP-IN／FRT-pcDNA5和ECMV-IRES的抗体双表达载体，把抗HBsAg抗体轻链、重链基因克隆到该表达栽体上进行表达。结果与结论：在CHO细胞中瞬时表达时没有检测到抗体表达，但在用抗性筛选到的克隆中有稳定的表达，各克隆之间表达量差异小于40％，为各种不同基因工程抗体表达量的筛选和优化提供了方法。

Bi-cistronic expression of anti-HBsAg based on Flp recombination targeting and IRES sequence

Objective: To compare the expression effi ci ency of whole antibodies that are different in construction. Methods: A bi-cistronic antibody expression vector was constructed that inclu ded FLP recombination site and ECMV-IRES sequence.The light chain was inserted up-streamly to the IRES site, and in down-stream multi-cloning site, the he av y chain was inserted. The construct was transfected into CHO/dhfr- cell with L ipofectAMINE.Results and Conclusion: In transient expressi on, no detectableantibody expression was produced in ELISA assay, but in stable clone selected with blamycin, the antibody was detected, and the expre ss ion variance was less than 40%.It was proved that the construct is a promising t ool for protein expression level comparing.