Antibody Phage Display pp 155-163

Part of the Methods in Molecular Biology book series (MIMB, volume 562)

Selection of Antibodies Able to Rapidly Enter Mammalian Cells

Protocol

Summary

This chapter outlines a protocol for the selection by phage display of single-chain variable antibody fragments with dual properties-specificity for tumor cells and the ability to be internalized. The protocol is based on a direct incubation of living target cells with antibody phage display libraries under conditions that allow active endocytosis of phage particles by cancer cells as well as recovery of intracellular phage particles that retain infectivity. This “functional” selection helps avoid the isolation of irrelevant phages that may be obtained when selection is performed on heterogeneous material as a source of antigen. Internalizing antibodies recovered from human antibody repertoires are promising reagents for various therapeutic applications including the delivery of drugs to the cytosol of targeted cell populations.

Key words:

Single-chain variable antibody fragment Cancer cell targeting Internalizing antibody 

References

  1. 1.
    Nielsen, U. B., and Marks, J. D. (2000) Internalizing antibodies and targeted cancer therapy: direct selection from phage display libraries. Pharm. Sci. Technol. Today 3, 282–291PubMedCrossRefGoogle Scholar
  2. 2.
    Becerril, B., Poul, M. A., and Marks, J. D. (1999) Toward selection of internalizing antibodies from phage libraries. Biochem. Biophys. Res. Commun. 255, 386–393PubMedCrossRefGoogle Scholar
  3. 3.
    O’Connell, D., Becerril, B., Roy-Burman, A., Daws, M., and Marks, J. D. (2002) Phage versus phagemid libraries for generation of human monoclonal antibodies. J. Mol. Biol. 321, 49–56PubMedCrossRefGoogle Scholar
  4. 4.
    Poul, M. A., Becerril, B., Nielsen, U. B., Morisson, P., and Marks, J. D. (2000) Selection of tumor-specific internalizing human antibodies from phage libraries. J. Mol. Biol. 301, 1149–1161PubMedCrossRefGoogle Scholar
  5. 5.
    Mazuet, C., Lerouge, D., Poul, M. A., and Blin, N. (2006) Breast carcinoma specific antibody selection combining phage display and immunomagnetic cell sorting. Biochem. Biophys. Res. Commun. 348, 550–559PubMedCrossRefGoogle Scholar
  6. 6.
    Liu, B., Conrad, F., Cooperberg, M. R., Kirpotin, D. B., and Marks, J. D. (2004) Mapping tumor epitope space by direct selection of single-chain Fv antibody libraries on prostate cancer cells. Cancer Res. 64, 704–710PubMedCrossRefGoogle Scholar
  7. 7.
    An, F., Drummond, D. C., Wilson, S., Kirpotin, D. B., Nishimura, S. L., Broaddus, V. C., and Liu, B. (2008) Targeted drug delivery to mesothelioma cells using functionally selected internalizing human single-chain antibodies. Mol. Cancer Ther. 7, 569–578PubMedCrossRefGoogle Scholar
  8. 8.
    Schier, R., Marks, J. D., Wolf, E. J., Apeldl, G., Wong, C., McCartney, J. E., Bookman, M. A., Huston, J. S., Houston, L. L., Weiner, L. M., and Adams, G. P. (1995) In vitro and in vivo characterization of a human anti-c-erbB-2 single-chain Fv isolated from a filamentous phage antibody library. Immunotechnology 1, 73–81PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Laboratoire de Biotechnologie et Pharmacologie Génétique Appliquée (LBPA)UMR CNRS 8113Ecole Normale Supérieure de CachanFrance

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