Antibody phage display libraries have frequently been used for the isolation of antibody fragments against potential target structures on the cell surface of different types of cancer. Independent from the source of V-regions and the antibody format, two general strategies have been followed in generating antibody phage display libraries. Either non-immune so-called naïve (or synthetic) libraries or repertoires from immunized animals (or patients) were generated. Especially antibody phage display libraries derived from immunized mice represent an attractive approach because many classical immunization protocols are available and have been successfully used in combination with phage display. Using these techniques, valuable high affinity antibody fragments have been isolated and served as tools in research as well as starting points for the development of antibody-based therapeutics. In this chapter two strategies to generate scFv fragments that recognize surface antigens in their native states by using mouse immune libraries are described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chowdhury PS, Viner JL, Beers R, Pastan I (1998) Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity. Proc Natl Acad Sci USA 95:669–674
Eylar E, Hagopian A (1971) Isolation of plasma membranes from mammalian cells. In: Jakoby WB (ed) Methods in enzymology, vol XXII. Academic, London
Groves M, Lane S, Douthwaite J, Lowne D, Rees DG, Edwards B, Jackson RH (2006) Affinity maturation of phage display antibody populations using ribosome display. J Immunol Methods 313:129–139
Harlow E, Lane D (1988) Antibodies – A laboratory manual. Cold Spring Harbor Laboratory Press, New York
Hoogenboom HR (2005) Selecting and screening recombinant antibody libraries. Nat Biotechnol 23:1105–1116
Hwang WY, Foote J (2005) Immunogenicity of engineered antibodies. Methods 36:3–10
Knappik A, Ge L, Honegger A, Pack P, Fischer M, Wellnhofer G, Hoess A, Wolle J, Pluckthun A, Virnekas B (2000) Fully synthetic human combinatorial antibody libraries (HuCAL), based on modular consensus frameworks and CDRs randomized with trinucleotides. J Mol Biol 296:57–86
Krebber A, Bornhauser S, Burmester J, Honegger A, Willuda J, Bosshard HR, Pluckthun A (1997) Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system. J Immunol Methods 201:35–55
Nissim A, Hoogenboom HR, Tomlinson IM, Flynn G, Midgley C, Lane D, Winter G (1994) Antibody fragments from a ‘single pot' phage display library as immunochemical reagents. EMBO J 13:692–698
Osbourn J, Groves M, Vaughan T (2005) From rodent reagents to human therapeutics using antibody guided selection. Methods 36:61–68
Peipp M, Simon N, Loichinger A, Baum W, Mahr K, Zunino SJ, Fey GH (2001) An improved procedure for the generation of recombinant single-chain Fv antibody fragments reacting with human CD13 on intact cells. J Immunol Methods 251:161–176
Schwemmlein M, Peipp M, Barbin K, Saul D, Stockmeyer B, Repp R, Birkmann J, Oduncu F, Emmerich B, Fey GH (2006) A CD33-specific single-chain immunotoxin mediates potent apoptosis of cultured human myeloid leukaemia cells. Br J Haematol 133:141–151
Thie H, Meyer T, Schirrmann T, Hust M, Dübel S (2008) Phage display derived therapeutic antibodies. Curr Pharm Biotechnol 9:439–446
Winter G, Griffiths AD, Hawkins RE, Hoogenboom HR (1994) Making antibodies by phage display technology. Annu Rev Immunol 12:433–455
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
Kellner, C., Nodehi, S.M., Peipp, M. (2010). Mouse Immune Libraries for the Generation of ScFv Fragments Directed Against Human Cell Surface Antigens. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01144-3_4
Download citation
DOI: https://doi.org/10.1007/978-3-642-01144-3_4
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-01143-6
Online ISBN: 978-3-642-01144-3
eBook Packages: Springer Protocols