medigraphic.com
ISSN 1027-2852 (Electronic)
ISSN 0864-4551 (Print)

2013, Number 1

<< Back Next >>

Biotecnol Apl 2013; 30 (1)

Replacement of serum supplemented medium for CB.Hep-1 hybridoma cell freezing and monoclonal antibody production

Aragón H, González M, Valdés R, Álvarez T, Brown E, Rodríguez Y, González T, Somoza R, Wood M

Language: English
References: 28
Page: 57-62
PDF size: 215.91 Kb.


ABSTRACT

CB.Hep-1 monoclonal antibody (mAb) has been used in Hepatitis B vaccine manufacture. The mAb production was performed by ascites method inoculating cells previously frozen and cultivated in fetal bovine serum supplemented medium (SSM). It is expensive at risk and can be contaminated with adventitious pathogens. Therefore, the main objective of this study was to replace the SSM by the chemically defi ned medium Filocethplus (cell freezing) and the protein free medium HyQ®SFM4CHO™ (cell growing). As results, cells cryo-preserved in Filocethplus for 180 days and cultivated in HyQ®SFM4CHO™ showed no significant differences in regards to viability, maximum viable cell density and growth conditions with the same cells cultivated and frozen in SSM. In addition, CB.Hep-1 cells that were frozen in Filocethplus and cultivated in HyQ®SFM4CHO™ exhibited similar performance to those frozen and cultivated in SSM in agitated culture, which point out the application of these media for CB.Hep-1 mAb and Hepatitis B vaccine production.


REFERENCES

  1. Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefi ned specifi city. Nature. 1975; 256(5517):495-7.

  2. Cobo F, Talavera P, Concha A. Diagnostic approaches for viruses and prions in stem cell banks. Virology. 2006;347(1): 1-10.

  3. Even MS, Sandusky CB, Barnard ND. Serum-free hybridoma culture: ethical, scientifi c and safety considerations. Trends Biotechnol. 2006;24(3):105-8.

  4. Zhang X, Stettler M, Reif O, Kocourek A, Dejesus M, Hacker DL, et al. Shaken helical track bioreactors: Providing oxygen to high-density cultures of mammalian cells at volumes up to 1000 L by surface aeration with air. N Biotechnol. 2008;25(1):68-75.

  5. Wurm FM. Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotechnol. 2004; 22(11):1393-8.

  6. Kim DY, Lee JC, Chang HN, Oh DJ. Development of serum-free media for a recombinant CHO cell line producing recombinant antibody. Enzyme Microb Technol. 2006;39(3):426-33.

  7. Schroder M, Matischak K, Friedl P. Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11. J Biotechnol. 2004;108(3): 279-92.

  8. Chen Z, Ke Y, Chen Y. A serum-free medium for hybridoma cell culture. Cytotechnology. 1993;11(3):169-74.

  9. Glassy MC, Tharakan JP, Chau PC. Serum-free media in hybridoma culture and monoclonal antibody production. Biotechnol Bioeng. 1988;32(8):1015-28.

  10. Martinelle K, Westlund A, Haggstrom L. Ammonium ion transport-a cause of cell death. Cytotechnology. 1996;22(1-3): 251-4.

  11. Goergen JL, Marc A, Engasser JM. Determination of cell lysis and death kinetics in continuous hybridoma cultures from the measurement of lactate dehydrogenase release. Cytotechnology. 1993;11(3):189-95.

  12. Thermo Scientifi c- HyClone- SFM4- CHO- Cell Culture Media Thermo Scientifi c* HyClone* SFM4CHO* Cell Culture Media [Internet]. Waltham, MA: Thermo Fisher Scientific Inc.; c2012 [cited 2012 Feb 12]. Thermo Fisher Scientifi c Inc.; [about 1 screen]. Available from: http://www.fi shersci.com/ecomm/servlet/ Search?keyWord=Thermo+Scientific- +HyClone-+SFM4CHO-+Cell+Culture+ Media&store=&nav=0&offSet=0&storeId =10652&langId=-1&searchType=PROD

  13. González Y, Fischer RW. Serum-free culturing of mammalian cells-adaptation to and cryopreservation in fully defi ned media. ALTEX. 2007;24(2):110-6.

  14. Fontirrochi G, Dueñas M, Fernández de Cossío ME, Fuentes P, Pérez M, Mainet D, et al. A mouse hybridoma cell line secreting IgG and IgM antibodies with specifi city for the hepatitis B surface antigen. Biotecnol Apl. 1993;10(1):24-30.

  15. Patterson MK, Jr. Measurement of growth and viability of cells in culture. Methods Enzymol. 1979;58:141-52.

  16. Valdés R, Álvarez T, Tamayo A, Fernández EG, Montero J, Geada D, et al. New Mab CB.Hep-1 Purifi cation Process Eliminates the Need for Pre-Chromatographic Purifi cation. Stability Demonstrated Over 100 Purifi cation Cycles. Chromatographia. 2008;67(11-12):923-7.

  17. Harshman JS. Purifi cation of murine monoclonal antibodies by affi nity chromatography using protein A-Sepharose. Methods Cell Sci. 2005;9(3):183-6.

  18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75.

  19. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5.

  20. Shinmoto H, Dosako S. Adaptation of mouse-human hybridomas to a protein free medium. Biotechnol Lett. 1993;15(4): 327-30.

  21. Darfl er FJ. A protein-free medium for the growth of hybridomas and other cells of the immune system. In Vitro Cell Dev Biol. 1990;26(8):769-78.

  22. Liguori MJ, Hoff-Velk JA, Ostrow DH. Recombinant human interleukin-6 enhances the immunoglobulin secretion of a rabbit-rabbit hybridoma. Hybridoma. 2001;20(3):189-98.

  23. Schneider YJ. Optimisation of hybridoma cell growth and monoclonal antibody secretion in a chemically defi ned, serumand protein-free culture medium. J Immunol Methods. 1989;116(1):65-77.

  24. Patchornik G, Albeck A. A general approach for antibody purifi cation utilizing [Protein A-catechol:Fe3+] macrocomplexes. Bioconjugate Chem. 2006; 17(2):258-60.

  25. Hahn R, Shimahara K, Steindl F, Jungbauer A. Comparison of protein A affi nity sorbents III. Life time study. J Chromatogr A. 2006;1102(1-2):224-31.

  26. Hober S, Nord K, Linhult M. Protein A chromatography for antibody purifi cation. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;848(1):40-7.

  27. Hahn R, Schlegel R, Jungbauer A. Comparison of protein A affi nity sorbents. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;790(1-2):35-51.

  28. Tavares RM, Karmali A, Clemente A, Pais MS. Production and characterization of a specifi c rubisco monoclonal antibody, and its use in rubisco quantifi cation during Zantedeschia aethiopica spathe development. Hybridoma. 1999;18(2):203-9.




2020     |     www.medigraphic.com