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Construction of Chicken Antibody Libraries

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Phage Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

Recombinant antibody libraries based on chicken immunoglobulin genes are potentially valuable sources of phage displayed scFvs for use in veterinary diagnostics and research. The libraries described in this chapter are based on chicken variable heavy and light chain immunoglobulin genes joined by a short flexible linker cloned in the phagemid vector pHEN1. The resulting phagemids produce either scFvs displayed on the surface of the fusion phage subsequent to co-infection with helper phage, or soluble scFvs following IPTG induction. This chapter provides detailed and proven methods for the construction of such libraries.

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References

  1. Davies EL, Smith JS, Birkett CR, Manser JM, Anderson-Dear DV, Young JR (1995) Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes. J Immunol Methods 186:125–135

    Article  CAS  PubMed  Google Scholar 

  2. Yamanaka HI, Inoue T, Ikeda-Tanaka O (1996) Chicken monoclonal antibody isolated by a phage display system. J Immunol 157:1156–1162

    CAS  PubMed  Google Scholar 

  3. Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF III (2000) Methods for the generation of chicken monoclonal antibody fragments by phage display. J Immunol Methods 242:159–181

    Article  CAS  PubMed  Google Scholar 

  4. Abi-Ghanem D, Waghela SD, David J, Caldwell DJ, Danforth HD, Berghman LR (2008) Phage display selection and characterization of single-chain recombinant antibodies against Eimeria tenella sporozoites. Vet Immunol Immunopathol 121:58–67

    Article  CAS  PubMed  Google Scholar 

  5. Chiliza TE, Van Wyngaardt W, Du Plessis DH (2008) Single-chain antibody fragments from a display library derived from chickens immunized with a mixture of parasite and viral antigens. Hybridoma 27:412–421. https://doi.org/10.1089/hyb.2008.0051

    Article  Google Scholar 

  6. Abolnik CA, Fehrsen J, Olivier A, van Wyngaardt W, Fosgate G, Ellis C (2013) Serological investigation of highly pathogenic avian influenza (HPAI) H5N2 in ostriches (Struthio camelus). Avian Pathol. https://doi.org/10.1080/03079457.2013.779637

  7. Li B, Yea J, Lin Y, Wanga M, Zhua J (2014) Preparation and identification of a single-chain variable fragment antibody against Newcastle diseases virus F48E9. Vet Immunol Immunopathol 161:258–264

    Article  CAS  PubMed  Google Scholar 

  8. Huston JS, Levinson D, Mudgett-Hunter M, Tai M-S, Novotný J, Margolies MN, Ridge RJ, Bruccoleri RE, Haber E, Crea R, Oppermann H (1988) Protein engineering of antibody binding sites: Recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc Natl Acad Sci U S A 85:5879–5883

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Hoogenboom HR, Griffiths AD, Johnson KS, David J, 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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. 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

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Van Wyngaardt W, Malatji T, Mashau C, Fehrsen J, Jordaan F, Miltiadou DR, Du Plessis DH (2004) A large semi-synthetic single-chain Fv phage display library based on chicken immunoglobulin genes. BMC Biotechnol 4:6

    Article  PubMed  PubMed Central  Google Scholar 

  12. Sixholo J, van Wyngaardt W, Mashau C, Frischmuth J, Du Plessis DH, Fehrsen J (2011) Improving the characteristics of a mycobacterial 16kDa-specific chicken scFv. Biologicals 39:110–116

    Article  CAS  PubMed  Google Scholar 

  13. Clackson T, Hoogenboom HR, Griffiths A, Winter G (1991) Making antibody fragments using phage display libraries. Nature 352:624–628

    Article  CAS  PubMed  Google Scholar 

  14. Fehrsen J, van Wyngaardt W, Mashau C, Potgieter C, Chaudhary VK, Gupta A, Jordaan F, du Plessis DH (2005) Serogroup-reactive and type-specific detection of bluetongue virus antibodies using chicken scFvs in inhibition ELISAs. J Virol Methods 129(1):31–39

    Article  CAS  PubMed  Google Scholar 

  15. Wemmer S, Mashau C, Fehrsen J, Wyngaardt W, du Plessis DH (2010) Chicken scFvs and bivalent scFv-CH fusions directed against HSP65 of Mycobacterium bovis. Biologicals 38:407–414

    Article  CAS  PubMed  Google Scholar 

  16. Rakabe M, Van Wyngaardt W, Fehrsen J (2011) Chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus. Food Agric Immunol 22(3):p283–p295

    Article  Google Scholar 

  17. Opperman PA, Maree FF, Van Wyngaardt W, Vosloo W, Theron J (2012) Mapping of antigenic determinants on a SAT2 foot-and-mouth disease virus using chicken single-chain antibody fragments. Virus Res 167:370–379

    Article  CAS  PubMed  Google Scholar 

  18. Van Wyngaardt W, Mashau C, Wright I, Fehrsen J (2013) Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs. J Vet Sci 14:95–98

    Article  PubMed  PubMed Central  Google Scholar 

  19. Atrazhef AM, Elliot JF (1996) Simplified desalting of ligation reactions immediately prior to electroporation into E. coli. Biotechniques 21:1024

    Google Scholar 

  20. Sambrook J, Russel DW (eds) (2001) Molecular cloning. A laboratory manual, Standard ethanol precipitation of DNA in microfuge tubes, vol 3, 3rd edn. Cold Spring Harbour Laboratory Press, Cold Spring Harbour, NY, p A8.14

    Google Scholar 

  21. Clackson T, Lowman HB (eds) (2004) Phage display. A practical approach. Oxford University Press, New York, NY

    Google Scholar 

  22. Barbas CF III, Burton DR, Scott JK, Silverman GJ (eds) (2001) Phage display. A laboratory manual. Cold Spring Harbour Laboratory Press, Cold Spring Harbour, NY

    Google Scholar 

  23. Rand KN (1996) Crystal violet can be used to visualize DNA bands during gel electrophoresis and to improve cloning efficiency. Trends J Tech Tips Online 1:23–24

    Article  Google Scholar 

  24. Schade R, Staak C, Hendriksen C, Erhard M, Hugl H, Koch G, Larsson A, Pollmann W, van Regenmortel M, Rijke E, Spielmann H, Steinbusch SD (1996) The production of avian (egg yolk) antibodies: IgY. ATLA 24:925–934

    Google Scholar 

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Acknowledgments

We thank Dr. Dion H du Plessis, our now retired research leader and mentor who started the phage display group. We are grateful to Dr. Marco Romito for the veterinary support, the good care of our chickens over the years and even taking them out for walks. The Medical Research Council (Cambridge, UK) for the gift of pHENI vector. Funders that made the work possible include Agricultural Research Council-Onderstepoort Veterinary Research, Innovation Fund of the Department of Science and Technology, and National Department of Agriculture both of South Africa.

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Authors and Affiliations

  1. Immunology Division, New Generation Vaccines Programme, ARC-Onderstepoort Veterinary Research, Onderstepoort, Pretoria, 0110, South Africa

    Jeanni Fehrsen, Susan Wemmer & Wouter van Wyngaardt

Authors
  1. Jeanni Fehrsen
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  2. Susan Wemmer
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  3. Wouter van Wyngaardt
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Corresponding author

Correspondence to Jeanni Fehrsen .

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Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Fehrsen, J., Wemmer, S., van Wyngaardt, W. (2018). Construction of Chicken Antibody Libraries. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_10

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

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