Abstract
Traditional methods of phage display panning bind purified antigen to plates or other solid phases to which libraries are then applied. These methods are not directly applicable to antigens in their native environment on cell surfaces or in settings where the target antigen is unknown. We describe here a procedure of a panning strategy on cell surface receptors including a depletion step. We explain every step of the protocol: production of phage library, depletion and selection, elution, screening by ELISA, and analysis of soluble antibodies by ELISA and flow cytometry. Finally, several possible variants of the protocol are explained in Subheading 4.
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
McCafferty J, Griffiths AD, Winter G, Chiswell DJ (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554
Fuchs P, Breitling F, Dubel S, Seehaus T, Little M (1991) Targeting recombinant antibodies to the surface of Escherichia coli: fusion to a peptidoglycan associated lipoprotein. Biotechnology (N Y) 9:1369–1372
Zahnd C, Amstutz P, Pluckthun A (2007) Ribosome display: selecting and evolving proteins in vitro that specifically bind to a target. Nat Methods 4:269–279
Boder ET, Wittrup KD (1997) Yeast surface display for screening combinatorial polypeptide libraries. Nat Biotechnol 15:553–557
Andersen PS, Stryhn A, Hansen BE, Fugger L, Engberg J, Buus S (1996) A recombinant antibody with the antigen-specific, major histocompatibility complex-restricted specificity of T cells. Proc Natl Acad Sci U S A 93:1820–1824
Barry MA, Dower WJ, Johnston SA (1996) Toward cell-targeting gene therapy vectors: selection of cell-binding peptides from random peptide-presenting phage libraries. Nat Med 2:299–305
Cai X, Garen A (1995) Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells: selection of specific antibodies from single-chain Fv fusion phage libraries. Proc Natl Acad Sci U S A 92:6537–6541
de Kruif J, Terstappen L, Boel E, Logtenberg T (1995) Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library. Proc Natl Acad Sci U S A 92:3938–3942
Marks JD, Ouwehand WH, Bye JM, Finnern R, Gorick BD, Voak D, Thorpe SJ, Hughes-Jones NC, Winter G (1993) Human antibody fragments specific for human blood group antigens from a phage display library. Biotechnology (N Y) 11:1145–1149
Hart SL, Knight AM, Harbottle RP, Mistry A, Hunger HD, Cutler DF, Williamson R, Coutelle C (1994) Cell binding and internalization by filamentous phage displaying a cyclic Arg-ÂGly-Asp-containing peptide. J Biol Chem 269:12468–12474
Poul MA, Becerril B, Nielsen UB, Morisson P, Marks JD (2000) Selection of tumor-specific internalizing human antibodies from phage libraries. J Mol Biol 301:1149–1161
Meulemans EV, Nieland LJ, Debie WH, Ramaekers FC, van Eys GJ (1995) Phage displayed antibodies specific for a cytoskeletal antigen. Selection by competitive elution with a monoclonal antibody. Hum Antibodies Hybridomas 6:113–118
Li J, Lundberg E, Vernet E, Larsson B, Hoiden-Guthenberg I, Graslund T (2010) Selection of affibody molecules to the ligand-binding site of the insulin-like growth factor-1 receptor. Biotechnol Appl Biochem 55:99–109
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Even-Desrumeaux, K., Chames, P. (2012). Phage Display and Selections on Cells. In: Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 907. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-974-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-61779-974-7_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-973-0
Online ISBN: 978-1-61779-974-7
eBook Packages: Springer Protocols