Bioprocess and Biosystems Engineering

, Volume 41, Issue 7, pp 961–971 | Cite as

All-trans retinoic acid in combination with sodium butyrate enhances specific monoclonal antibody productivity in recombinant CHO cell line

  • Mahmood Rahimi-Zarchi
  • Seyed Abbas Shojaosadati
  • Mohammad Mehdi Amiri
  • Mahmood Jeddi-Tehrani
  • Fazel Shokri
Research Paper


The effects of all-trans retinoic acid (RA) and sodium butyrate (NaBu) on growth, viability and antibody production of two types of transfected Chinese hamster ovary cell lines (CHO-K1 and CHO-S) were investigated using a batch mode cell culture. By adding 0.5 mM NaBu in the CHO-K1 cell culture, the cell specific productivity (Qp) and antibody concentration increased by five- and threefold, respectively. The optimal concentration of RA was 100 nM which resulted in twofold increase in antibody production. In a combination model, RA applied at early growth phase of CHO-K1 cells followed by addition of NaBu with lowering culture temperature at the end of stationary phase resulted in two- and threefold increase in Qp and final antibody concentration, respectively. The latter strategy was also applied on suspended CHO-S cells with enhanced Qp and antibody concentration, but to a lesser extent than the CHO-K1 cells. In conclusion, our results demonstrate that the addition of RA and NaBu along with lowering the culture temperature can increase cell culture period as well as Qp and the final concentration of recombinant monoclonal antibody in both CHO-K1 and CHO-S cells without any significant change in binding affinity of the mAb.


CHO cells Monoclonal antibody Sodium butyrate All-trans retinoic acid Cell specific productivity (Qp



This work was supported partially by grants from Tarbiat Modares University and Tehran University of Medical Sciences.

Compliance with ethical standards

Conflict of interest

We declare that there is no conflict of interest.


  1. 1.
    Walsh G (2014) Biopharmaceutical benchmarks 2014. Nat Biotechnol 32(10):992–1000. CrossRefPubMedGoogle Scholar
  2. 2.
    Konno Y, Aoki M, Takagishi M, Sakai N, Koike M, Wakamatsu K, Hosoi S (2011) Enhancement of antibody production by the addition of Coenzyme-Q(10). Cytotechnology 63(2):163–170. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Choi SS, Rhee Wj Fau - Kim EJ, Kim Ej Fau -. Park TH, Park TH (2006) Enhancement of recombinant protein production in Chinese hamster ovary cells through anti-apoptosis engineering using 30Kc6 gene. Biotechnol Bioeng 95(3):459–467. CrossRefPubMedGoogle Scholar
  4. 4.
    Chen K, Liu Q, Xie L, Sharp PA, Wang DI (2001) Engineering of a mammalian cell line for reduction of lactate formation and high monoclonal antibody production. Biotechnol Bioeng 72 (1):55–61.<55::AID-BIT8>3.0.CO;2-4CrossRefPubMedGoogle Scholar
  5. 5.
    Kim DY, Lee JF, Chang HN, Chang Hn Fau - Oh DJ, Oh DJ (2005) Effects of supplementation of various medium components on chinese hamster ovary cell cultures producing recombinant antibody. In: Cytotechnology (0920–9069 (Print)):37–49. doi:D-NLM: PMC3449820 EDAT-2008/11/13 09:00 MHDA-2008/11/13 09:01 CRDT-2008/11/13 09:00 PHST-2005/03/31 [received] PHST-2005/07/29 [accepted] AID— PST-publish
  6. 6.
    Sun Y-t, Zhao L, Ye Z, Fan L, Liu X-p, Tan W-S (2013) Development of a fed-batch cultivation for antibody-producing cells based on combined feeding strategy of glucose and galactose. Biochem Eng J 81:126–135. CrossRefGoogle Scholar
  7. 7.
    Kumar N, Gammell P, Clynes M (2007) Proliferation control strategies to improve productivity and survival during CHO based production culture: a summary of recent methods employed and the effects of proliferation control in product secreting CHO cell lines. Cytotechnology 53(1–3):33–46. CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Sunley K, Butler M (2010) Strategies for the enhancement of recombinant protein production from mammalian cells by growth arrest. Biotechnol Adv 28(3):385–394. CrossRefPubMedGoogle Scholar
  9. 9.
    Park JH, Noh SM, Woo JR, Kim JW, Lee GM (2016) Valeric acid induces cell cycle arrest at G1 phase in CHO cell cultures and improves recombinant antibody productivity. Biotechnol J 11(4):487–496. CrossRefPubMedGoogle Scholar
  10. 10.
    Ahn WS, Jeon J-J, Jeong Y-R, Lee SJ, Yoon SK (2008) Effect of culture temperature on erythropoietin production and glycosylation in a perfusion culture of recombinant CHO cells. Biotechnol Bioeng 101(6):1234–1244. CrossRefPubMedGoogle Scholar
  11. 11.
    Chen Z-L, Wu B-C, Liu H, Liu X-M, Huang P-T (2004) Temperature shift as a process optimization step for the production of pro-urokinase by a recombinant Chinese hamster ovary cell line in high-density perfusion culture. J Biosci Bioeng 97(4):239–243. CrossRefPubMedGoogle Scholar
  12. 12.
    Allen MJ, Boyce JP, Trentalange MT, Treiber DL, Rasmussen B, Tillotson B, Davis R, Reddy P (2008) Identification of novel small molecule enhancers of protein production by cultured mammalian cells. Biotechnol Bioeng 100(6):1193–1204. CrossRefPubMedGoogle Scholar
  13. 13.
    Bi J-X, Shuttleworth J, Al-Rubeai M (2004) Uncoupling of cell growth and proliferation results in enhancement of productivity in p21CIP1-arrested CHO cells. Biotechnol Bioeng 85(7):741–749. CrossRefPubMedGoogle Scholar
  14. 14.
    Wurm MF (2013) CHO quasispecies—implications for manufacturing processes. Processes. CrossRefGoogle Scholar
  15. 15.
    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. CrossRefPubMedGoogle Scholar
  16. 16.
    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. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Hong JK, Lee SM, Kim K-Y, Lee GM (2014) Effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant Chinese hamster ovary cells. Appl Microbiol Biotechnol 98(12):5417–5425. CrossRefPubMedGoogle Scholar
  18. 18.
    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. CrossRefPubMedGoogle Scholar
  19. 19.
    Chen F, Kou T, Fan L, Zhou Y, Ye Z, Zhao L, Tan W-S (2011) The combined effect of sodium butyrate and low culture temperature on the production, sialylation, and biological activity of an antibody produced in CHO cells. Biotechnol Bioprocess Eng 16(6):1157–1165. CrossRefGoogle Scholar
  20. 20.
    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. CrossRefPubMedGoogle Scholar
  21. 21.
    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. CrossRefPubMedGoogle Scholar
  22. 22.
    Cherlet M, Marc A (2000) Stimulation of monoclonal antibody production of hybridoma cells by butyrate: evaluation of a feeding strategy and characterization of cell behaviour. Cytotechnology 32(1):17–29. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Sung YH, Lee JS, Park SH, Koo J, Lee GM (2007) Influence of co-down-regulation of caspase-3 and caspase-7 by siRNAs on sodium butyrate-induced apoptotic cell death of Chinese hamster ovary cells producing thrombopoietin. Metab Eng 9(5–6):452–464. CrossRefPubMedGoogle Scholar
  24. 24.
    Oh HK, So MK, Yang J, Yoon HC, Ahn JS, Lee JM, Kim JT, Yoo JU, Byun TH (2005) Effect of N-acetylcystein on butyrate-treated chinese hamster ovary cells to improve the production of recombinant human interferon-β-1a. Biotechnol Prog 21(4):1154–1164. CrossRefPubMedGoogle Scholar
  25. 25.
    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. CrossRefPubMedGoogle Scholar
  26. 26.
    Kim NS, Lee GM (2000) Overexpression of bcl-2 inhibits sodium butyrate-induced apoptosis in Chinese hamster ovary cells resulting in enhanced humanized antibody production. Biotechnol Bioeng 71 (3):184–193.<184::AID-BIT1008>3.0.CO;2-WCrossRefPubMedGoogle Scholar
  27. 27.
    Sung YH, Hwang SF - Lee GM, Lee GM Influence of down-regulation of caspase-3 by siRNAs on sodium-butyrate-induced apoptotic cell death of Chinese hamster ovary cells producing thrombopoietin (1096–7176 (Print))Google Scholar
  28. 28.
    Inoue Y, Fujisawa M, Shoji M, Hashizume S, Katakura Y, Shirahata S (2000) Enhanced antibody production of human–human hybridomas by retinoic acid. Cytotechnology 33(1–3):83–88. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Inoue Y, Fujisawa M, Kawamoto S, Shoji M, Hashizume S, Fujii M, Katakura Y, Shirahata S (1999) Effectiveness of vitamin A acetate for enhancing the production of lung cancer specific monoclonal antibodies. Cytotechnology 31(1–2):77–83. CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Amiri MM, Jeddi-Tehrani M, Kazemi T, Bahadori M, Maddah M, Hojjat-Farsangi M, Khoshnoodi J, Rabbani H, Shokri F (2013) Construction and characterization of a new chimeric antibody against HER2. Immunotherapy 5(7):703–715. CrossRefPubMedGoogle Scholar
  31. 31.
    Tahmasebi F, Kazemi T, Amiri MM, Khoshnoodi J, Mahmoudian J, Bayat AA, Jeddi-Tehrani M, Rabbani H, Shokri F (2014) In vitro assessment of the effects of anti-HER2 monoclonal antibodies on proliferation of HER2-overexpressing breast cancer cells. Immunotherapy 6(1):(1750–7448 (Electronic)):43–49. CrossRefGoogle Scholar
  32. 32.
    Kazemi T, Tahmasebi F, Bayat AA, Mohajer N, Khoshnoodi J, Jeddi-Tehrani M, Rabbani H, Shokri F (2011) Characterization of novel murine monoclonal antibodies directed against the extracellular domain of human HER2 tyrosine kinase receptor. Hybridoma 30(4):347–353. CrossRefPubMedGoogle Scholar
  33. 33.
    Hajighasemi FS-YA., Shokri F (2004) Measurement of affinity constant of anti-human IgG monoclonal antibodies by an ELISA-based method. Irn J Immunol 1(3):154–161Google Scholar
  34. 34.
    Golsaz Shirazi F, Mohammadi H, Fau-Amiri MM, Singethan K, Xia Y, Bayat AA, Bahadori M, Rabbani H, Jeddi-Tehrani M, Protzer U, Shokri F (2014) Monoclonal antibodies to various epitopes of hepatitis B surface antigen inhibit hepatitis B virus infection. J Gastroenterol Hepatol 29(5):1083–1091. CrossRefPubMedGoogle Scholar
  35. 35.
    Du Z, Treiber D, McCarter JD, Fomina-Yadlin D, Saleem RA, McCoy RE, Zhang Y, Tharmalingam T, Leith M, Follstad BD, Dell B, Grisim B, Zupke C, Heath C, Morris AE, Reddy P (2015) Use of a small molecule cell cycle inhibitor to control cell growth and improve specific productivity and product quality of recombinant proteins in CHO cell cultures. Biotechnol Bioeng 112(1):141–155. CrossRefPubMedGoogle Scholar
  36. 36.
    Rahimpour A, Ahani R, Najaei A, Adeli A, Barkhordari F, Mahboudi F (2016) Development of genetically modified chinese hamster ovary host cells for the enhancement of recombinant tissue plasminogen activator expression. The Malaysian Journal of Medical Sciences: MJMS 23(2):6–13PubMedGoogle Scholar
  37. 37.
    Sharow KA, Temkin B, Asson-Batres MA (2012) Retinoic acid stability in stem cell cultures. Int J Dev Biol 56(4):273–278. CrossRefPubMedGoogle Scholar
  38. 38.
    Jefferis R (2016) Posttranslational Modifications and the Immunogenicity of Biotherapeutics. J Immunol Res 2016:5358272.

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mahmood Rahimi-Zarchi
    • 1
  • Seyed Abbas Shojaosadati
    • 1
  • Mohammad Mehdi Amiri
    • 2
  • Mahmood Jeddi-Tehrani
    • 3
  • Fazel Shokri
    • 2
  1. 1.Biotechnology Group, Faculty of Chemical EngineeringTarbiat Modares UniversityTehranIran
  2. 2.Department of Immunology, School of Public HealthTehran University of Medical SciencesTehranIran
  3. 3.Monoclonal Antibody Research CenterAvicenna Research Institute, ACECRTehranIran

Personalised recommendations