Hyperphage is a substitute for M13KO7 helper phage used in antibody phage display. It allows to improve the antibody display efficiency on phage by 2-3 orders of magnitude. This is achieved by forcing the packaging E. coli cell to exclusively use the pIII portion of the antibody fusion protein for its virion assembly. Hyperphage is compatible with all usual phagemid systems using a full length pIII gene in fusion to the binder repertoire, and has been widely validated to improve panning with antibody and scaffold libraries.
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
Hust M, Dübel S, Schirrmann T (2007) Selection of recombinant antibodies from antibody gene libraries. In: Ochs M (ed) Gene function analysis. Humana Press, Totowa, NY, pp 243–255 ISBN 1597455474
Rondot S, Koch J, Breitling F, Dübel S (2001) A helper phage to improve single-chain antibody presentation in phage display. Nat Biotechnol 19:75–78
Soltes G, Hust M, Ng KKY, Bansal A, Field J, Stewart DIH, Dübel S, Cha S, Wiersma E (2007) On the influence of vector design on antibody phage display. J Biotechnol 127:626–637
Barbas CF III, Kang AK, Lerner RA, Benkovic SJ (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci USA 88:7978–7982
Breitling F, Dübel S, Seehaus T, Klewinghaus I, Little M (1991) A surface expression vector antibody screening. Gene 104:147–153
Hoogenboom HR, Griffith AD, Johnson KS, Chiswell DJ, Hudson P, Winter G (1991) Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucleic Acids Res 19:4133–4137
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbour Laboratory Press, New York
Koch J, Breitling F, Dübel S (2000) Rapid titration of multiple samples of filamentous bacteriophage (M13) on nitrocellulose filters. Biotechniques 29:1196–1202
Micheel B, Heymann S, Scharte G, Böttger V, Vogel F, Dübel S, Breitling F, Little M, Behrsing O (1994) Production of monoclonal antibodies against epitopes of the main coat protein of filamentous fd phage. J Immunol Methods 171(1):103–109
Taussig MJ, Stoevesandt O, Borrebaeck C, Bradbury A, Dübel S, Frank R, Gibson T, Gold L, Herberg F, Hermjakob H, Hoheisel J, Joos T, Konthur Z, Landegren U, Plückthun A, Ueffing M, Uhlén M (2007) ProteomeBinders: planning a European resource of affinity reagents for analysis of the human proteome. Nat Methods 4:13–17
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer-Verlag
About this protocol
Cite this protocol
Breitling, F., Broders, O., Helmsing, S., Hust, M., Dübel, S. (2010). Improving Phage Display Throughput by Using Hyperphage, Miniaturized Titration and pVIII (g8p) ELISA. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01144-3_14
Download citation
DOI: https://doi.org/10.1007/978-3-642-01144-3_14
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-01143-6
Online ISBN: 978-3-642-01144-3
eBook Packages: Springer Protocols