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12 (2) 2022

A preliminary study to establish the transfected CHO cell lines which highly express Trastuzumab - A biosimilar product of Herceptin


Author - Affiliation:
Nguyen Quang Huy - University of Science, Vietnam National University - HCMC Navetco J.S.C, Ho Chi Minh City , Vietnam
Huynh Thi Yen Ly - Nanogen J.S.C, Ho Chi Minh City , Vietnam
Tran Thi Nhu Mai - Nanogen J.S.C, Ho Chi Minh City , Vietnam
Nguyen Thi Thanh Truc - Nanogen J.S.C, Ho Chi Minh City , Vietnam
Huynh Trong Hieu - Nanogen J.S.C, Ho Chi Minh City
Nguyen Mai Khoi - Nanogen J.S.C, Ho Chi Minh City
Doan Chinh Chung - Nanogen J.S.C, Ho Chi Minh City
Trinh Thanh Hung - Ministry of Science and Technology, Ho Chi Minh City
Do Minh Si - University of Science, Vietnam National University - HCMC Nanogen J.S.C, Ho Chi Minh City
Corresponding author: Nguyen Quang Huy - nguyenquanghuy166@gmail.com
Submitted: 24-01-2022
Accepted: 25-03-2022
Published: 04-11-2022

Abstract
Human epidermal growth factor receptor 2 (HER2) has been identified as a molecular target for breast cancer therapy, such as Trastuzumab (Herceptin®). This has been shown to improve patient survival substantially. The current study is aiming to locally produce an anti-HER2 monoclonal antibody (named Trastuzumab) which has an equivalent biological properties in comparison with the original version, Herceptin®). In silico design and construction of recombinant vectors, as well as the establishment of transfected cell lines with high expression of Trastuzumab were performed. Based on the protein sequences obtained from the Drugbank, the DNA sequences encoding for the light chain (Tras-Lc) and heavy chain (Tras-Hc) of Trastuzumab were optimized and integrated into pNanogen-Hygro and pNanogen-Puro vectors, respectively. The Neon Transfection System was used to co-transfect the pNanogen-Tras-Lc-Hygro and pNanogen-Tras-Hc-Puro constructs into CHO cells. Different co-transfected single-cell-colonies selected on media supplemented with hygromycin and puromycin were used for ELISA and SDS-PAGE assays to identify the CHO cell lines which highly express Trastuzumab. Based on the present results, 30μg of both constructs were suitable for DNA co-transfection. After 07 days of culture, the highest amount of Trastuzumab (561 µg/ml) was obtained from the H06LD68 cell line.

Keywords
CHO cell line; pNanogen; trastuzumab

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Cite this paper as:

Nguyen, H. Q., Huynh, L. T. Y., Tran, M. T. N., Nguyen, T. T. T., Huynh, H. T., Nguyen, K. M., Doan, C. C., Trinh, H. T., & Do, S. M. (2022). A preliminary study to establish the transfected CHO cell lines which highly express Trastuzumab - A biosimilar product of Herceptin. Ho Chi Minh City Open University Journal of Science – Engineering and Technology, 12(2), 60-69. doi:10.46223/HCMCOUJS.tech.en.12.2.2172.2022


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DOI: https://doi.org/10.46223/HCMCOUJS.tech.en.12.2.2172.2022
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