High-yield CHO-K1 instantaneous transfection method with linearized PEI as transfection reagent
By optimizing the transient transfection method of CHO-K1 cells, using linearized PEI and herring sperm DNA as alternative expression vectors, and combining GS expression vectors with temperature control, the problem of low transfection efficiency of CHO-K1 cells was solved, achieving efficient and economical recombinant protein production, which is suitable for bioreactor scale-up.
Patent Information
- Application Number
- CN202511573688.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-31
- Publication Date
- 2026-02-06
AI Technical Summary
The low transient transfection efficiency of CHO-K1 cells limits the production efficiency of recombinant protein drugs. The application of existing linearized PEI transfection reagents in CHO-K1 cells has not been systematically optimized, and traditional methods are time-consuming, making it difficult to meet the needs of rapid evaluation of candidate drugs.
Linearized PEI was used as the transfection reagent. The transfection process was optimized by completely replacing the culture medium and mixing it with DNA immediately, combined with herring sperm DNA replacement expression vector and GS expression vector. This included gradient temperature control and feeding strategy, which shortened the operation time and improved the transfection efficiency and expression level.
It significantly improves the transfection efficiency of CHO-K1 cells, with antibody expression levels exceeding 1 g/L, reduces transfection reagent costs, shortens the R&D cycle, ensures batch-to-batch consistency of the process, and is suitable for scale-up to bioreactor production.