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Effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant Chinese hamster ovary cells

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Abstract

Sodium butyrate (NaBu) is known to increase the specific productivity of recombinant Chinese hamster ovary (rCHO) cells. To understand the effects of NaBu on the product quality, rCHO cells producing monoclonal antibody (Mab) were cultivated at various concentrations of NaBu (0 to 4 mM). NaBu increased correctly assembled Mab. In the absence of NaBu, the proportions of intact Mab (2H2L) and heavy chain dimer (2H) were 81 and 15 %. At 1 mM NaBu, the proportion of 2H2L increased to 93 %, whereas the proportion of 2H decreased to 2 %. No further increase in the proportion of 2H2L was obtained at a higher NaBu concentration. NaBu also affected the charge heterogeneity of Mab, which may affect the efficacy of Mab. The basic charge variants of Mabs increased with an increase in the NaBu concentration. In addition, NaBu affected the galactosylation of Mab negatively. Overall, the data obtained here show that NaBu used in rCHO cell cultures for improved Mab production affects certain quality aspects of Mab, in this case, the charge heterogeneity and galactosylation.

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References

  • Bergman LW, Harris E, Kuehl WM (1981) Glycosylation causes an apparent block in translation of immunoglobulin heavy chain. J Biol Chem 256(2):701–716

    CAS  PubMed  Google Scholar 

  • Boswell CA, Tesar DB, Mukhyala K, Theil FP, Fielder PJ, Khawli LA (2010) Effects of charge on antibody tissue distribution and pharmacokinetics. Bioconjug Chem 21(12):2153–2163

    Article  CAS  PubMed  Google Scholar 

  • Brekke OH, Sandlie I (2003) Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 2:52–62

    Article  CAS  PubMed  Google Scholar 

  • Chang MC, Tsai YL, Chen YW, Chan CP, Huang CF, Lan WC, Lin CC, Lan WH, Jeng JH (2013) Butyrate induces reactive oxygen species production and affects cell cycle progression in human gingival fibroblasts. J Periodontal Res 48(1):66–73

    Article  CAS  PubMed  Google Scholar 

  • Chotigeat W, Watanapokasin Y, Mahler S, Gray PP (1994) Role of environmental conditions on the expression levels, glycoform pattern and levels of sialyltransferase for hFSH produced by recombinant CHO cells. Cytotechnology 15(1–3):217–221

    Article  CAS  PubMed  Google Scholar 

  • Chumsae C, Gaza-Bulseco G, Sun J, Liu H (2007) Comparison of methionine oxidation in thermal stability and chemically stressed samples of a fully human monoclonal antibody. J Chromatogr B Analyt Technol Biomed Life Sci 850(1–2):285–294

    Article  CAS  PubMed  Google Scholar 

  • Harris RJ, Kabakoff B, Macchi FD, Shen FJ, Kwong M, Andya JD, Shire SJ, Bjork N, Totpal K, Chen AB (2001) Identification of multiple sources of charge heterogeneity in a recombinant antibody. J Chromatogr B 752(2):233–245

    Article  CAS  Google Scholar 

  • Harris RJ, Shire SJ, Winter C (2004) Commercial manufacturing scale formulation and analytical characterization of therapeutic recombinant antibodies. Drug Dev Res 61(3):137–154

    Article  CAS  Google Scholar 

  • Hendrick V, Winnepenninckx P, Abdelkafi C, Vandeputte O, Cherlet M, Marique T, Renemann G, Loa A, Kretzmer G, Werenne J (2001) Increased productivity of recombinant tissular plasminogen activator (t-PA) by butyrate and shift of temperature: a cell cycle phases analysis. Cytotechnology 36(1–3):71–83

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hong JK, Lee GM, Yoon SK (2011) Growth factor withdrawal in combination with sodium butyrate addition extends culture longevity and enhances antibody production in CHO cells. J Biotechnol 155(2):225–231

    Article  CAS  PubMed  Google Scholar 

  • Hossler P, Khattak SF, Li ZJ (2009) Optimal and consistent protein glycosylation in mammalian cell culture. Glycobiology 19(9):936–949

    Article  CAS  PubMed  Google Scholar 

  • Hwang SO, Lee GM (2008) Nutrient deprivation induces autophagy as well as apoptosis in Chinese hamster ovary cell culture. Biotechnol Bioeng 99(3):678–685

    Article  CAS  PubMed  Google Scholar 

  • Jefferis R (2009) Glycosylation as a strategy to improve antibody-based therapeutics. Nat Rev Drug Discov 8:226–234

    Article  CAS  PubMed  Google Scholar 

  • Jiang Z, Sharfstein ST (2008) Sodium butyrate stimulates monoclonal antibody over-expression in CHO cells by improving gene accessibility. Biotechnol Bioeng 100(1):189–194

    Article  CAS  PubMed  Google Scholar 

  • Kaschak T, Boyd D, Lu F, Derfus G, Kluck B, Nogal B, Emery C, Summers C, Zheng K, Bayer R, Amanullah A, Yan BX (2011) Characterization of the basic charge variants of a human IgG1: effect of copper concentration in cell culture media. Mabs-Austin 3(6):577–583

    Article  Google Scholar 

  • Khawli LA, Goswami S, Hutchinson R, Kwong ZW, Yang JH, Wang XD, Yao ZL, Sreedhara A, Cano T, Tesar D, Nijem I, Allison DE, Wong PY, Kao YH, Quan C, Joshi A, Harris RJ, Motchnik P (2010) Charge variants in IgG1 Isolation, characterization, in vitro binding properties and pharmacokinetics in rats. Mabs-Austin 2(6):613–624

    Article  Google Scholar 

  • Kim NS, Lee GM (2002) Inhibition of sodium butyrate-induced apoptosis in recombinant Chinese hamster ovary cells by constitutively expressing antisense RNA of caspase-3. Biotechnol Bioeng 78(2):217–228

    Article  CAS  PubMed  Google Scholar 

  • Kim SJ, Kim NS, Ryu CJ, Hong HJ, Lee GM (1998) Characterization of chimeric antibody producing CHO cells in the course of dihydrofolate reductase-mediated gene amplification and their stability in the absence of selective pressure. Biotechnol Bioeng 58(1):73–84

    Article  CAS  PubMed  Google Scholar 

  • Lee JS, Lee GM (2012) Effect of sodium butyrate on autophagy and apoptosis in Chinese hamster ovary cells. Biotechnol Prog 28:349–357

    Article  CAS  PubMed  Google Scholar 

  • Lee SM, Kim YG, Lee EG, Lee GM (2014) Digital mRNA profiling of N-glycosylation gene expression in recombinant Chinese hamster ovary cells treated with sodium butyrate. J Biotechnol 171:56–60

    Article  CAS  PubMed  Google Scholar 

  • Leitzgen K, Knittler MR, Haas IG (1997) Assembly of immunoglobulin light chains as a prerequisite for secretion. A model for oligomerization-dependent subunit folding. J Biol Chem 272(5):3117–3123

    Article  CAS  PubMed  Google Scholar 

  • Lin HY, Massowelch P, Di YP, Cai JW, Shen JW, Subjeck JR (1993) The 170-kDa glucose-regulated stress protein is an endoplasmic reticulum protein that binds immunoglobulin. Mol Biol Cell 4(11):1109–1119

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Liu H, Gaza-Bulseco G, Faldu D, Chumsae C, Sun J (2008) Heterogeneity of monoclonal antibodies. J Pharm Sci 97(7):2426–2447

    Article  CAS  PubMed  Google Scholar 

  • Louis M, Rosato RR, Brault L, Osbild S, Battaglia E, Yang XH, Grant S, Bagrel D (2004) The histone deacetylase inhibitor sodium butyrate induces breast cancer cell apoptosis through diverse cytotoxic actions including glutathione depletion and oxidative stress. Int J Oncol 25(6):1701–1711

    CAS  PubMed  Google Scholar 

  • Malhotra JD, Miao H, Zhang K, Wolfson A, Pennathur S, Pipe SW, Kaufman RJ (2008) Antioxidants reduce endoplasmic reticulum stress and improve protein secretion. Proc Natl Acad Sci U S A 105(47):18525–18530

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Melnick J, Dul JL, Argon Y (1994) Sequential interaction of the chaperones BiP and GRP94 with immunoglobulin chains in the endoplasmic reticulum. Nature 370(6488):373–375

    Article  CAS  PubMed  Google Scholar 

  • Mimura Y, Lund J, Church S, Dong S, Li J, Goodall M, Jefferis R (2001) Butyrate increases production of human chimeric IgG in CHO-K1 cells whilst maintaining function and glycoform profile. J Immunol Methods 247(1–2):205–216

    Article  CAS  PubMed  Google Scholar 

  • Pan H, Chen K, Chu L, Kinderman F, Apostol I, Huang G (2009) Methionine oxidation in human IgG2 Fc decreases binding affinities to protein A and FcRn. Protein Sci 18(2):424–433

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Reddy P, Sparvoli A, Fagioli C, Fassina G, Sitia R (1996) Formation of reversible disulfide bonds with the protein matrix of the endoplasmic reticulum correlates with the retention of unassembled Ig light chains. EMBO J 15(9):2077–2085

    CAS  PubMed Central  PubMed  Google Scholar 

  • Rodriguez J, Spearman M, Huzel N, Butler M (2005) Enhanced production of monomeric interferon-beta by CHO cells through the control of culture conditions. Biotechnol Prog 21(1):22–30

    Article  CAS  PubMed  Google Scholar 

  • Santell L, Ryll T, Etcheverry T, Santoris M, Dutina G, Wang A, Gunson J, Warner TG (1999) Aberrant metabolic sialylation of recombinant proteins expressed in Chinese hamster ovary cells in high productivity cultures. Biochem Biophys Res Commun 258(1):132–137

    Article  CAS  PubMed  Google Scholar 

  • Schlatter S, Stansfield SH, Dinnis DM, Racher AJ, Birch JR, James DC (2005) On the optimal ratio of heavy to light chain genes for efficient recombinant antibody production by CHO cells. Biotechnol Prog 21(1):122–133

    Article  CAS  PubMed  Google Scholar 

  • Shapiro AL, Scharff MD, Maizel JV, Uhr JW (1966) Synthesis of excess light chains of gamma globulin by rabbit lymph node cells. Nature 211(5046):243–245

    Article  CAS  PubMed  Google Scholar 

  • Sung YH, Lee GM (2005) Enhanced human thrombopoietin production by sodium butyrate addition to serum-free suspension culture of bcl-2-overexpressing CHO cells. Biotechnol Prog 21(1):50–57

    Article  CAS  PubMed  Google 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

    Article  CAS  PubMed  Google Scholar 

  • Vanhoutvin SA, Troost FJ, Hamer HM, Lindsey PJ, Koek GH, Jonkers DM, Kodde A, Venema K, Brummer RJ (2009) Butyrate-induced transcriptional changes in human colonic mucosa. PLoS One 4(8):e6759

    Article  PubMed Central  PubMed  Google Scholar 

  • Vlasak J, Ionescu R (2008) Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. Curr Pharm Biotechnol 9(6):468–481

    Article  CAS  PubMed  Google Scholar 

  • Yoon SK, Hong JK, Lee GM (2004) Effect of simultaneous application of stressful culture conditions on specific productivity and heterogeneity of erythropoietin in Chinese hamster ovary cells. Biotechnol Prog 20(4):1293–1296

    Article  CAS  PubMed  Google Scholar 

  • Zhang T, Bourret J, Cano T (2011) Isolation and characterization of therapeutic antibody charge variants using cation exchange displacement chromatography. J Chromatogr A 1218(31):5079–5086

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported in part by the Converging Research Center Program through the NRF funded by the MEST (2009–0082276) and a grant from the Fundamental R&D Program for Technology of World Premier Materials funded by the Ministry of Knowledge Economy, Republic of Korea.

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Authors and Affiliations

  1. Department of Biological Sciences, KAIST, 335 Gwahak-ro, Yuseong-gu, Daejeon, 305-701, South Korea

    Jong Kwang Hong, Sang Min Lee & Gyun Min Lee

  2. ISU ABXIS, Global R&D Center, 696-1 Sampyeong-dong, Bundang-gu, Seongnam, 463-400, South Korea

    Kyung-Yong Kim

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  1. Jong Kwang Hong
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  2. Sang Min Lee
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  3. Kyung-Yong Kim
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  4. Gyun Min Lee
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Correspondence to Gyun Min Lee.

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Hong, J.K., Lee, S.M., Kim, KY. et al. Effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant Chinese hamster ovary cells. Appl Microbiol Biotechnol 98, 5417–5425 (2014). https://doi.org/10.1007/s00253-014-5596-8

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