Generation of CHO-DG44 cells producing recombinant anti-TNFα mAb with high productivity. Evaluation of purified anti-TNFα mAb quality in comparison with innovator drug, adalimumab.

Theoretical results:
- Construction of pN-ADA-LC/HC vectors to express mAbTNFα in CHO cell line.
- Generation of mAbTNFα-producing CHO cells with productivity > 0.5 g/L.
- Establishment of cell bank (CHO-ADA) and in-house specification.
- Optimization of cell culture condition on CHO-ADA with productivity > 2.5 g/L under optimized conditions in pilot-scale (5L) culture.
- Establishment of mAbTNFα purification process with efficiency > 60%, purity > 90%.
- Structural and biological characterization of mAbTNFα in comparison with adalimumab

Applied results: The findings of the project can provide an important fundament for the use of CHO host cells and designed vector system in the prodution of therapeutic proteins, especially monoclonal antibodies. Furthermore, The process described, in this study, can be applied as an effective approach for enhancing therapeutic antibody production in large-scale.

- The combination of UCOEs and DHFR in expression vectors is reported to significantly increase the expression of transgenes (HC or LC) in CHO cells.
- The use of UCOE and DHFR vectors in combination with the optimization of cell culture media may help in establishing stable and high-producing CHO cell lines for therapeutic antibody production.

- Scientific papers in referred journals
+ Chinh Chung Doan, Nguyen Quynh Chi Ho, Thi Thuy Nguyen, Thi Phuong Thao Nguyen, Dang Giap Do, Nghia Son Hoang, Thanh Long Le. Enhancement of anti-TNFα monoclonal antibody production in CHO cells through the use of UCOE and DHFR elements in vector construction and the optimization of cell culture media. Preparative Biochemistry & Biotechnology. 2021, 24: 1-19; DOI: 10.1080/10826068.2021.1963981.
+ Chinh Chung Doan, Thanh Long Le, Nguyen Quynh Chi Ho, Nghia Son Hoang. Effects of ubiquitous chromatin opening element (UCOE) on recombinant anti-TNFα antibody production and expression stability in CHO-DG44 cells. Cytotechnology. 2022, 74: 31-49; DOI: 10.1007/s10616-021-00503-1.
- Technological products (describe in details: technical characteristics, place)
+ pN-ADA-LC and pN-ADA-HC vectors: The UCOE and DHFR-based vectors containing the codon-optimized genes encoding HC and LC of mAbTNFα expressed in CHO cells.
+ CHO-ADA (mAbTNFα-producing CHO cells): The antibody concentration of CHO-ADA cultured in non-optimized conditions was approximately 0.5 g/L
+ mAbTNFα (fully humanized monoclonal anti-TNFα antibodies): The primary chemical and biological characteristics of mAbTNFα were similar to those of adalimumab
+ These technological products were deposited in Department of Animal Biotechnology, Institute of Tropical Biology
- Other products (if applicable)
+ Protocol for construction of vector expressing HC or LC of mAbTNFα in CHO cells.
+ Protocol for generation of mAbTNFα-producing CHO cells.
+ Protocol for purification of mAbTNFα.
+ Report for quality assessment of purified mAbTNFα in comparison with adalimumab.