Abstract
Monoclonal antibodies are increasingly used for treatment of various diseases, mostly cancer therapy and auto-immune diseases. New chimeric, humanized, or fully-human antibodies specific to novel or already validated targets are being developed in many academic labs as well as in commercial entities. A validation of antibodies in preclinical settings is an important component of the road from discovery to clinic. Preparing sufficient amount of high-quality antibodies for preclinical studies is a challenge in some cases. This article summarizes protocols used in research laboratories, and is based on our own experience in the establishment of stable clones of CHO cells to scaling-up of tissue cultures for production of antibodies in bioreactors.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
Abbreviations
- Ab:
-
antibody
- IgG:
-
immunoglobulin
- Fab:
-
antigen-binding fragment
- CDR:
-
complementarity-determining regions
- CHO:
-
Chinese hamster ovary
- HRP:
-
horseradish peroxidase
References
Johansson, D. X., Drakenberg, K., Hopmann, K. H., Schmidt, A., Yari, F., Hinkula, J., and Persson, M. A. (2007) Efficient expression of recombinant human monoclonal antibodies in Drosophila S2 cells. J. Immunol. Methods 318:37–46.
Zhu, L., van de Lavoir, M. C., Albanese, J., Beenhouwer, D. O., Cardarelli, P. M., Cuison, S., Deng, D. F., Deshpande, S., Diamond, J. H., Green, L., Halk, E. L., Heyer, B. S., Kay, R. M., Kerchner, A., Leighton, P. A., Mather, C. M., Morrison, S. L., Nikolov, Z. L., Passmore, D. B., Pradas-Monne, A., Preston, B. T., Rangan, V. S., Shi, M., Srinivasan, M., White, S. G., Winters-Digiacinto, P., Wong, S., Zhou, W., and Etches, R. J. (2005) Production of human monoclonal antibody in eggs of chimeric chickens. Nat. Biotechnol. 23:1159–69.
Werner, R. G., Noe, W., Kopp, K., and Schluter, M. (1998) Appropriate mammalian expression systems for biopharmaceuticals. Arzneimittelforschung 48:870–80.
Bender, E., Woof, J. M., Atkin, J. D., Barker, M. D., Bebbington, C. R., and Burton, D. R. (1993) Recombinant human antibodies: linkage of a Fab fragment from a combinatorial library to an Fc fragment for expression in mammalian cell culture. Hum. Antibodies Hybridomas 4:74–9.
Li, J., Menzel, C., Meier, D., Zhang, C., Dubel, S., and Jostock, T. (2007) A comparative study of different vector designs for the mammalian expression of recombinant IgG antibodies. J. Immunol. Methods 318:113–24.
Burton, D. R., Pyati, J., Koduri, R., Sharp, S. J., Thornton, G. B., Parren, P. W., Sawyer, L. S., Hendry, R. M., Dunlop, N., Nara, P. L., et al. (1994) Efficient neutralization of primary isolates of HIV-1 by a recombinant human monoclonal antibody. Science 266:1024–7.
Zhang, M. X., Bohlman, M. C., Itatani, C., Burton, D. R., Parren, P. W., St Jeor, S. C., and Kozel, T. R. (2006) Human recombinant antimannan immunoglobulin G1 antibody confers resistance to hematogenously disseminated candidiasis in mice. Infect. Immun. 74:362–9.
Wurm, F. M. (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat. Biotechnol. 22:1393–8.
Remaut, K., Sanders, N. N., Fayazpour, F., Demeester, J., and De Smedt, S. C. (2006) Influence of plasmid DNA topology on the transfection properties of DOTAP/DOPE lipoplexes. J. Control Release 115:335–43.
Acknowledgments
This research was supported by the Intramural research Program of the NIH, NCI, Center for Cancer Research.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Feng, Y., Dimitrov, D.S. (2009). Scaling-Up and Production of Therapeutic Antibodies for Preclinical Studies. In: Dimitrov, A. (eds) Therapeutic Antibodies. Methods in Molecular Biology™, vol 525. Humana Press. https://doi.org/10.1007/978-1-59745-554-1_26
Download citation
DOI: https://doi.org/10.1007/978-1-59745-554-1_26
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-934115-92-3
Online ISBN: 978-1-59745-554-1
eBook Packages: Springer Protocols