Related Experiment Video
Updated: May 9, 2026
Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety
Published on: September 15, 2023
The Role of Different Supplements in Expression Level of Monoclonal Antibody against Human CD20
Fereidoun Mahboudi1, Mohammad Reza Abolhassan, Armita Azarpanah
1Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Background:
Recombinant monoclonal antibodies have been marketed in last three decades as the major therapeutic proteins against different cancers. However choosing a proper medium and supplements to reach the high expression is a challenging step. Despite of commercial serum free and chemically defined media, there are still numerous researches seeking the optimum media to gain higher expression titer. Selecting the best basal media followed by proper supplementation to increase the cell density and expression titer needs proper and accurate investigation.
Methods:
In this study, we have determined the expression titer of monoclonal antibody against human CD20 using soy extract, Essential Amino Acid, Non-Essential Amino Acid, Panexin NTS, Peptone, Yeast extract, Insulin-transferrin selenite, Human Serum Albumin, Bovine Serum Albumin, Lipid, and two commercially available supplements, Power and Xtreme feed. In each experiment, the expression level was compared with a well defined media, ProCHO5, RPMI 1640 and DMEM-F12.
Results:
It has been shown that supplementing the ProCHO5 basal medium with 10% power feed or combination of 5% PanexinNTS,1.5 g/L yeast and 1.5g/L peptone results in the best production levels with 450 and 425 mg/L of anti CD20 mAb expression level, respectively.
Conclusion:
Panexin NTS, yeast and peptone cane be proper supplement for fed-batch cell culture instead of commercial Power feed supplement which is a cost effective way to increase expression level. And thereby ProCHO5 may be replaced with common media such as RPMI 1640 and DMEM-F12.
Insights
Optimizing cell culture media is crucial for high monoclonal antibody (mAb) expression. Supplementing ProCHO5 with yeast extract and peptone offers a cost-effective alternative to commercial feeds for enhanced anti-CD20 mAb production.
Area of Science:
- Biotechnology
- Biopharmaceutical Manufacturing
- Cell Culture Technology
Background:
- Recombinant monoclonal antibodies (mAbs) are key cancer therapeutics.
- Optimizing cell culture media and supplements for high mAb expression remains a challenge.
- Research continues to seek optimal media formulations for increased expression titers.
Purpose of the Study:
- To evaluate various supplements for enhancing monoclonal antibody expression.
- To identify cost-effective alternatives to commercial supplements.
- To compare expression levels in different basal media.
Main Methods:
- Assessed anti-CD20 mAb expression titers using diverse supplements including soy extract, amino acids, Panexin NTS, peptone, yeast extract, and albumin.
- Compared expression levels against defined media like ProCHO5, RPMI 1640, and DMEM-F12.
- Utilized commercially available supplements Power feed and Xtreme feed as benchmarks.
Main Results:
- Supplementing ProCHO5 with 10% Power feed yielded 450 mg/L of anti-CD20 mAb.
- A combination of 5% Panexin NTS, 1.5 g/L yeast extract, and 1.5 g/L peptone achieved 425 mg/L anti-CD20 mAb expression.
- These results highlight effective supplementation strategies for mAb production.
Conclusions:
- Panexin NTS, yeast extract, and peptone serve as effective supplements for fed-batch cell culture.
- This combination presents a cost-effective alternative to commercial Power feed.
- ProCHO5 can potentially be replaced by common media like RPMI 1640 and DMEM-F12 with appropriate supplementation.
