Abstract
Cell fusion protocols that were developed by Kohler and Milstein in the mid-1970s and aimed at producing and characterization of mouse monoclonal antibodies (MAbs) remain the gold standard of hybridoma development. Despite tremendous progress in using MAbs in multiple research, diagnostic, and therapeutic areas, major experimental flaws in designing and carrying out hybridoma experimentation often result in the production of hybridomas exhibiting poor growth parameters and secreting low-specificity and low-affinity antibodies. This methodology chapter is built around the conventional hybridoma protocol, with a special emphasis on tissue culture and biochemical techniques aimed at producing truly monospecific and highly active mouse MAbs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kohler, G. and Milstein, C. (1976) Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur. J. Immunol. 6, 511–519.
He, Y., Honnen, W. J., Krachmarov, C. P., et al. (2002) Efficient isolation of novel human monoclonal antibodies with neutralizing activity against HIV-1 from transgenic mice expressing human Ig loci. J. Immunol. 169, 595–605.
Hoogenboom, H. R. and Chames, P. (2000) Natural and designer binding sites made by phage display technology. Immunol. Today 21, 371–378.
Liu, B., Huang, L., Sihlbom, C., Burlingame, A., and Marks, J. D. (2002) Towards proteome-wide production of monoclonal antibody by phage display. J. Mol. Biol. 315, 1063–1073.
Hanes, J., Schaffitzel, C., Knappik, A., and Pluckthun, A. (2000) Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display. Nat. Biotechnol. 18, 1287–1292.
Tomlinson, I. and Holliger, P. (2000) Methods for generating multivalent and bispecific antibody fragments. Methods Enzymol. 326, 461–479.
Proske, D., Blank, M., Buhmann, R., and Resch, A. (2005) Aptamers —basic research, drug development, and clinical applications. Appl. Microbiol. Biotechnol. 69, 367–374.
Hudson, P. J. and Souriau, C. Engineered antibodies. (2003) Nat. Med. 9, 129–134.
Jain, K. K. (2005) Personalised medicine for cancer: from drug development into clinical practice. Expert Opin. Pharmacother. 6, 1463–1476.
Bioinfomatics (2004) The market for antibodies: keys to success for commercial suppliers. Report #04-049.
deStGroth, S. F. and Scheidegger, D. (1980) Production of monoclonal antibodies: strategy and tactics. J. Immunol. Methods 35, 1–21.
Mechetner, E. and Roninson, I. (1998) Monoclonal antibody to a human MDR1 multidrug resistance gene product, and uses. US Patent no. 5,773,280.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Mechetner, E. (2007). Development and Characterization of Mouse Hybridomas. In: Albitar, M. (eds) Monoclonal Antibodies. Methods in Molecular Biology, vol 378. Humana Press. https://doi.org/10.1007/978-1-59745-323-3_1
Download citation
DOI: https://doi.org/10.1007/978-1-59745-323-3_1
Publisher Name: Humana Press
Print ISBN: 978-1-58829-567-5
Online ISBN: 978-1-59745-323-3
eBook Packages: Springer Protocols