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Updated: Jul 6, 2025
Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
Strategy for Developing a Stable CHO Cell Line that Produces Large Titers of Trastuzumab Antibody
Hafsa Boulenouar1,2, Nadia Bouchoutrouch2, Youssef Amar2
1Laboratory of Pharmacology & Toxicology, Faculty of Medicine & Pharmacy, University Mohammed V in rabat, 10170 Rabat, Morocco.
Background:
Trastuzumab (Herceptin®) is currently the main treatment option for breast cancer patients that overexpress the human epidermal growth factor receptor 2 (HER2). This antibody binds specifically to HER2, blocks cancer cell growth, and promotes effective cell death. In the present study, we sought to develop a robust and efficient process for the development of a stable Chinese hamster ovary (CHO) cell line with high trastuzumab expression and production.
Methods:
We adapted a process that combines transposon system-based vector construction, suspension cell culture, and a high selection process. The latter, involved enhanced green fluorescent protein (eGFP) expression, fluorescence-activated cell sorting (FACS), and semi-solid methylcellulose media.
Results:
The construction of trastuzumab as a humanized monoclonal antibody was achieved by subcloning the synthesized light and heavy chain sequences into a suitable piggyBac expression vector. The optimized piggyBac vector used for the expression of trastuzumab in CHO cells resulted in the production of trastuzumab and reached 4.24 g/L in the T1A7 clone after a 7-day batch culture. The T1A7 clone was selected after screening over 1500 clones.
Conclusions:
The current simple workflow ensures strict monoclonality and relatively high production of trastuzumab. This workflow could potentially be implemented in Research and Development (R&D) laboratories, including in developing countries for the production of recombinant monoclonal antibodies in a cost-effective manner.
Insights
Researchers developed a streamlined process for creating stable Chinese hamster ovary (CHO) cell lines that efficiently produce trastuzumab, a key antibody for HER2-positive breast cancer treatment. This method enhances antibody production and offers a cost-effective solution for research and development.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Trastuzumab (Herceptin®) is a primary treatment for HER2-overexpressing breast cancer.
- It targets HER2, inhibiting cancer cell growth and promoting cell death.
Purpose of the Study:
- To develop a robust and efficient process for creating a stable Chinese hamster ovary (CHO) cell line.
- The goal was high trastuzumab expression and production.
Main Methods:
- Utilized a transposon system for vector construction.
- Employed suspension cell culture with enhanced green fluorescent protein (eGFP) expression.
- Incorporated fluorescence-activated cell sorting (FACS) and semi-solid methylcellulose media for high selection.
Main Results:
- Successfully constructed trastuzumab as a humanized monoclonal antibody using a piggyBac expression vector.
- Achieved trastuzumab production of 4.24 g/L in the T1A7 clone after a 7-day batch culture.
- Selected the T1A7 clone from over 1500 screened clones.
Conclusions:
- The developed workflow ensures monoclonality and high trastuzumab production.
- This cost-effective method is suitable for R&D laboratories, including those in developing countries.
- Facilitates the production of recombinant monoclonal antibodies.
