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

Recently, there has been a high demand for anti-tumor necrosis factor-α monoclonal antibodies (mAbTNFα) in the treatment of rheumatoid arthritis and other autoimmune diseases. Thus, efficient strategies and stable high-producing cell lines need to be established to increase antibody production. In t...

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Bibliographic Details
Published in:Preparative biochemistry & biotechnology 2022, Vol.52 (4), p.452-470
Main Authors: Doan, Chinh Chung, Ho, Nguyen Quynh Chi, Nguyen, Thi Thuy, Nguyen, Thi Phuong Thao, Do, Dang Giap, Hoang, Nghia Son, Le, Thanh Long
Format: Article
Language:English
Subjects:
Adalimumab
Animals
anti-TNFα monoclonal antibody
Antibodies
Antibodies, Monoclonal - genetics
Antibody Formation
Arthritis
Autoimmune diseases
Batch culture
Cell culture
Cell Culture Techniques
CHO Cells
CHO-DG44 cell line
Chromatin
Cricetinae
Cricetulus
Culture media
DHFR
Dihydrofolate reductase
Dilution
Expression vectors
gene amplification
Media
medium optimization
Methotrexate
Monoclonal antibodies
Optimization
Reductases
Rheumatoid arthritis
Tetrahydrofolate Dehydrogenase - genetics
Tetrahydrofolate Dehydrogenase - metabolism
TNF inhibitors
Tumor Necrosis Factor-alpha - genetics
Tumor necrosis factor-α
UCOE element
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Summary:Recently, there has been a high demand for anti-tumor necrosis factor-α monoclonal antibodies (mAbTNFα) in the treatment of rheumatoid arthritis and other autoimmune diseases. Thus, efficient strategies and stable high-producing cell lines need to be established to increase antibody production. In this study, we describe an efficient approach to establish a mAbTNFα high-producing clone through the optimization of expression vectors and cell culture media. The ubiquitous chromatin opening element (UCOE) and dihydrofolate reductase (DHFR)-based vectors encoding mAbTNFα were introduced into the CHO-DG44 cells using lipofection. Clones were obtained by selecting transfected cells with G418, amplifying them by treatment with methotrexate, and isolating them by limiting dilution. Different media formulated with commercial feeds and media were also screened to develop an improved medium. The antibody produced by the selected clone was purified, characterized, and compared to standard adalimumab. Using our established protocol, a cell clone obtained from stable mAbTNFα-expressing cell pools showed a 3.8-fold higher antibody titer compared to stable cell pools. Furthermore, the highest antibody yield of selected clones cultured in fed-batch mode using improved medium was 2450 ± 30 µg/mL, which was 13.2-fold higher than that of stable cell pool cultivated in batch mode using a basal medium. The purified antibody had primary chemical and biological characteristics similar to those of adalimumab. Therefore, 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.
ISSN:1082-6068
1532-2297
DOI:10.1080/10826068.2021.1963981