Biphasic addition strategy of hypoxanthine and thymidine for improving monoclonal antibody production.
In our previous study, we demonstrated that combinatorial addition of hypoxanthine (10 mg/L) and thymidine (2 mg/L) was able to stimulate initial cell growth and elevate volumetric concentration of antibody by 22% (Chen et al., Appl. Microbiol. Biotechnol., 93, 169-178, 2012). In this study, a systematic study was carried out to investigate the effects of hypoxanthine and thymidine (H&T) on cell growth and antibody production in a much wider range of concentration. In addition, we pursued to establish a highly productive fed-batch culture via rationally designing H&T addition regime. It was found that both cell growth and antibody production in batch cultures were H&T concentration-dependent. Specifically, a low concentration stimulated cell growth while exerting no influence on specific productivity (q(mAb)), and a high concentration inhibited cell growth, however, significantly enhancing q(mAb). Subsequent experiments with fed-batch shaking flasks demonstrated the feasibility of improving antibody production using a biphasic addition strategy for H&T: supplementing a low concentration of H&T during initial cell growth phase and a high concentration of H&T at the production phase. By applying the optimized feeding regime, a maximum viable cell density (VCD) of 6.45 × 10(6)cells/mL and volumetric antibody production of 632 mg/L were achieved in a 2 L-B.Braun bioreactor. Taken together, in this study, a biphasic H&T addition strategy for cell culture was developed, which hold great promise to improve antibody production.
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, P.O. Box 309, Shanghai 200237, PR China.
Fei
F
Chen
Zhaoyang
Z
Ye
Liang
L
Zhao
Xuping
X
Liu
Li
L
Fan
Wen-Song
WS
Tan
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation / Q000187:drug effects; D061249:Batch Cell Culture Techniques / Q000379:methods; D019149:Bioreactors; D016466:CHO Cells; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D019271:Hypoxanthine / Q000378:metabolism / Q000494:pharmacology*; D013936:Thymidine / Q000378:metabolism / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D019271:Hypoxanthine; D013936:Thymidine