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. 2013 Jul;5(3):140-7.

The Role of Different Supplements in Expression Level of Monoclonal Antibody against Human CD20

Fereidoun Mahboudi  1 , Mohammad Reza Abolhassan, Armita Azarpanah, Hamideh Aghajani-Lazarjani, Mohammad Amin Sadeghi-Haskoo, Shaian Maleknia, Behrouz Vaziri
Affiliations

The Role of Different Supplements in Expression Level of Monoclonal Antibody against Human CD20

Fereidoun Mahboudi et al. Avicenna J Med Biotechnol. 2013 Jul.

Abstract

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.

Keywords: CHO cells; Gene expression; Monoclonal antibody.

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Figures

Figure 1
Figure 1
Expression of recombinant mAb against h-CD20 in DMEM-F12. Recombinant CHO cells have been grown in DMEM-F12 and fed with different supplements. 0.2×106 cells/well seeded in 6-Well plate and cultured for 7 days. Supplement combination: 1. Yeast (1.5 g/L)+Peptone(1.5 g/L)+NTS (5%) 2. Yeast (1.5 g/L)+Peptone(1.5 g/L)+Sigma supplement (0.1%) 3. Yeast (1.5 g/L)+Peptone(1.5 g/L)+ITS(0.1%) 4. Essential amino acid (1%)+ Non-essential amino acid (1%) 5. Essential amino acid (1%)+ Non-essential amino acid (1%)+Lipid (1%) 6. Human Serum Albumin (HSA)(3%)+Sigma supplement (0.5%) 7. Control (DMEM-F12)
Figure 2
Figure 2
Expression of recombinant mAb against h- CD20 in RPMI 1640. Recombinant CHO cells have been grown in RPMI 1640 and fed with different supplements. 0.2×106 cells/well seeded in 6-Well plate and cultured for 7 days. Supplements combination 1. Yeast (1.5 g/L)+Peptone (1.5 g/L) 2. Yeast (1.5 g/L)+Peptone (1.5 g/L)+NTS (5%) 3. Yeast (1.5 g/L)+Peptone (1.5 g/L)+Sigma Supplement (0.1%) 4. Human Serum Albumin (HSA) (3%)+ Sigma supplement (0.5%) 5. Control (RPMI 1640)
Figure 3
Figure 3
Expression of recombinant mAb against h- CD20 in ProCHO5. Recombinant CHO cells have been grown in ProCHO5 and fed with different supplements. 0.2×106 cells/well seeded in 6-Well plate and cultured for 7 days Supplements combination 1. Peptone (1.5 g/L) 2. Yeast (1.5 g/L)+Peptone (1.5 g/L) 3. Yeast (1.5 g/L)+Peptone (1.5 g/L)+NTS (5%) 4. Control (ProCHO5)
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