The effect of hyperosmolality application time on production, quality, and biopotency of monoclonal antibodies produced in CHO cell fed-batch and perfusion cultures
Hyperosmolality has been commonly investigated due to its effects on the production and quality characteristics of monoclonal antibodies (mAbs) produced in CHO cell fed-batch cultures. However, the application of hyperosmolality at different times and its effect on biopotency have seldom been researched, especially in perfusion culture. In our study, different degrees of hyperosmolality induced by sodium chloride were investigated in anti-IgE rCHO cell fed-batch cultures and anti-CD52 rCHO cell perfusion cultures during the initial and stable phases. The results showed that the initial hyperosmolality group (IHG) in fed-batch and early phase of perfusion cultures exhibited significant suppression of the viable cell density yet an enhancement in specific productivity, whereas the stable hyperosmolality group (SHG) achieved higher mAb production in both fed-batch and perfusion cultures. Additionally, the SHG produced less aggregates and acidic charge variants than IHG in fed-batch culture, which differed from perfusion cultures. However, the contents of non-glycosylation heavy chain (NGHC) and man5 were higher in SHG than in IHG in fed-batch cultures at plus 60 and 120 mOsm/kg, which was similar to perfusion cultures. Furthermore, the biopotency in the IHG was higher than in the SHG at plus 60 and 120 mOsm/kg in fed-batch cultures, which is similar to complement-dependent cytotoxicity (CDC) efficacy in perfusion cultures. The biopotency of all group was acceptable, except FI3. Thus, the study shows that hyperosmolality at a certain level could be beneficial for both mAb production, quality and biopotency, which could play an important role in process development for commercial production.
KeywordsChinese hamster ovary (CHO) Hyperosmolality Fed-batch Perfusion Biopotency
This project was sponsored by Shanghai Taiyin Biotech Co. Ltd. We thank everyone who helped with this research.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
- Bertrand V, Vogg S, Villiger TK, Stettler M, Broly H, Soos M, Morbidelli M (2018) Proteomic analysis of micro-scale bioreactors as scale-down model for a mAb producing CHO industrial fed-batch platform. J Biotechnol 279:27–36. https://doi.org/10.1016/j.jbiotec.2018.04.015 CrossRefPubMedGoogle Scholar
- Grilo AL, Mantalaris A (2018) The increasingly human and profitable monoclonal antibody market. Trends Biotechnol. https://doi.org/10.1016/j.tibtech.2018.05.014
- Heidemann R, Zhang C, Qi H, Larrick Rule J, Rozales C, Park S, Chuppa S, Ray M, Michaels J, Konstantinov K, Naveh D (2000) The use of peptones as medium additives for the production of a recombinant therapeutic protein in high density perfusion cultures of mammalian cells. Cytotechnology 32(2):157–167. https://doi.org/10.1023/a:1008196521213 CrossRefPubMedPubMedCentralGoogle Scholar
- Hintersteiner B, Lingg N, Zhang P, Woen S, Hoi KM, Stranner S, Wiederkum S, Mutschlechner O, Schuster M, Loibner H, Jungbauer A (2016) Charge heterogeneity: basic antibody charge variants with increased binding to fc receptors. mAbs 8(8):1548–1560. https://doi.org/10.1080/19420862.2016.1225642 CrossRefPubMedPubMedCentralGoogle Scholar
- Kanda Y, Yamada T, Mori K, Okazaki A, Inoue M, Kitajima-Miyama K, Kuni-Kamochi R, Nakano R, Yano K, Kakita S, Shitara K, Satoh M (2007) Comparison of biological activity among nonfucosylated therapeutic IgG1 antibodies with three different N-linked fc oligosaccharides: the high-mannose, hybrid, and complex types. Glycobilogy 17(1):104–118. https://doi.org/10.1093/glycob/cwl057 CrossRefGoogle Scholar
- Konno Y, Kobayashi Y, Takahashi K, Takahashi E, Sakae S, Wakitani M, Yamano K, Suzawa T, Yano K, Ohta T, Koike M, Wakamatsu K, Hosoi S (2012) Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity. Cytotechnology 64(3):249–265. https://doi.org/10.1007/s10616-011-9377-2 CrossRefPubMedGoogle Scholar
- Leblanc Y, Ramon C, Bihoreau N, Chevreux G (2017) Charge variants characterization of a monoclonal antibody by ion exchange chromatography coupled on-line to native mass spectrometry: case study after a long-term storage at +5 degrees C. J Chromatogr B 1048:130–139. https://doi.org/10.1016/j.jchromb.2017.02.017 CrossRefGoogle Scholar
- Nasseri SS, Ghaffari N, Braasch K, Jardon MA, Butler M, Kennard M, Gopaluni B, Piret JM (2014) Increased CHO cell fed-batch monoclonal antibody production using the autophagy inhibitor 3-MA or gradually increasing osmolality. Biochem Eng J 91:37–45. https://doi.org/10.1016/j.bej.2014.06.027 CrossRefGoogle Scholar
- Shen D, Kiehl TR, Khattak SF, Li ZJ, He A, Kayne PS, Patel V, Neuhaus IM, Sharfstein ST (2010) Transcriptomic responses to sodium chloride-induced osmotic stress: a study of industrial fed-batch CHO cell cultures. Biotechnol Prog 26(4):1104–1115. https://doi.org/10.1002/btpr.398 CrossRefPubMedGoogle Scholar
- Sung YH, Song YJ, Lim SW, Chung JY, Lee GM (2004) Effect of sodium butyrate on the production, heterogeneity and biological activity of human thrombopoietin by recombinant Chinese hamster ovary cells. J Biotechnol 112(3):323–335. https://doi.org/10.1016/j.jbiotec.2004.05.003 CrossRefPubMedGoogle Scholar
- Xie P, Niu H, Chen X, Zhang X, Miao S, Deng X, Liu X, Tan WS, Zhou Y, Fan L (2016) Elucidating the effects of pH shift on IgG1 monoclonal antibody acidic charge variant levels in Chinese hamster ovary cell cultures. Appl Microbiol Biotechnol 100(24):10343–10353. https://doi.org/10.1007/s00253-016-7749-4 CrossRefPubMedGoogle Scholar
- Zheng C, Zhuang C, Chen Y, Fu Q, Qian H, Wang Y, Qin J, Wu X, Qi N (2018a) Improved process robustness, product quality and biological efficacy of an anti-CD52 monoclonal antibody upon pH shift in Chinese hamster ovary cell perfusion culture. Process Biochem 65:123–129. https://doi.org/10.1016/j.procbio.2017.11.013 CrossRefGoogle Scholar
- Zheng C, Zhuang C, Qin J, Chen Y, Fu Q, Qian H, Wu T, Wang Y, Wu X, Qi N (2018b) Combination of temperature shift and hydrolysate addition regulates anti-IgE monoclonal antibody charge heterogeneity in Chinese hamster ovary cell fed-batch culture. Cytotechnology 70(4):1121–1129. https://doi.org/10.1007/s10616-018-0192-x CrossRefPubMedGoogle Scholar
- Zhou W, Chen C-C, Buckland B, Aunins J (1997) Fed-batch culture of recombinant NS0 myeloma cells with high monoclonal antibody production. Biotechnol Bioeng 55(5):783–792. https://doi.org/10.1002/(SICI)1097-0290(19970905)55:5<783::AID-BIT8>3.0.CO;2-7 CrossRefPubMedGoogle Scholar
- Zhuang C, Zheng C, Chen Y, Huang Z, Wang Y, Fu Q, Zeng C, Wu T, Yang L, Qi N (2017) Different fermentation processes produced variants of an anti-CD52 monoclonal antibody that have divergent in vitro and in vivo characteristics. Appl Microbiol Biotechnol 101(15):5997–6006. https://doi.org/10.1007/s00253-017-8312-7 CrossRefPubMedGoogle Scholar