Whole-Genome Phage Display Libraries: A Powerful Tool for Antigen Discovery

  • Elisa BeghettoEmail author
  • Nicola Gargano
Part of the Methods in Molecular Biology book series (MIMB, volume 2024)


In the last two decades, phage display technology has been used for investigating complex biological processes and isolating molecules of practical value in several applications. Bacteriophage lambda, representing a classical cloning and expression system, has also been exploited for generating display libraries of small peptides and protein domains. More recently, large cDNA and whole-genome lambda display libraries of human pathogens have been generated for the discovery of new antigens for biomedical applications. Here, we describe the construction of a whole-genome library of a common pathogen—Streptococcus pneumoniae—and the use of this library for the molecular dissection of the human B-cell response against bacterial infection and colonization.

Key words

Bacteriophage lambda Phage display library Genomic DNA display Streptococcus pneumoniae Antigen discovery 


  1. 1.
    Mikawa YG, Maruyama IN, Brenner S (1996) Surface display of proteins on bacteriophage lambda heads. J Mol Biol 262:21–30CrossRefGoogle Scholar
  2. 2.
    Sternberg N, Hoess RH (1995) Display of peptides and proteins on the surface of bacteriophage lambda. Proc Natl Acad Sci U S A 92:1609–1613CrossRefGoogle Scholar
  3. 3.
    Niwa M, Maruyama H, Fujimoto T, Dohi K, Maruyama IN (2000) Affinity selection of cDNA libraries by lambda phage surface display. Gene 256:229–236CrossRefGoogle Scholar
  4. 4.
    Nicastro J, Sheldon K, Slavcev RA (2014) Bacteriophage lambda display systems: developments and applications. Appl Microbiol Biotechnol 98(7):2853–2866CrossRefGoogle Scholar
  5. 5.
    Beghetto E, Pucci A, Minenkova O, Spadoni A, Bruno L, Buffolano W, Soldati D, Felici F, Gargano N (2001) Identification of a human immunodominant B-cell epitope within the GRA1 antigen of Toxoplasma gondii by phage display of cDNA libraries. Int J Parasitol 31:1659–1668CrossRefGoogle Scholar
  6. 6.
    Minenkova O, Pucci A, Pavoni E, De Tomassi A, Fortugno P, Gargano N, Cianfriglia M, Barca S, De Placido S, Martignetti A, Felici F, Cortese R, Monaci P (2003) Identification of tumor-associated antigens by screening phage-displayed human cDNA libraries with sera from tumor patients. Int J Cancer 106:534–544CrossRefGoogle Scholar
  7. 7.
    Parmley SF, Smith GP (1988) Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene 73:305–318CrossRefGoogle Scholar
  8. 8.
    Enshell-Seijffers D, Gershoni JM (2002) Phage display selection and analysis of Ab-binding epitopes. Curr Protoc Immunol Chapter 9:Unit 9.8PubMedGoogle Scholar
  9. 9.
    Beghetto E, Spadoni A, Buffolano W, Del Pezzo M, Minenkova O, Pavoni E, Pucci A, Cortese R, Felici F, Gargano N (2003) Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries. Int J Parasitol 33:163–173CrossRefGoogle Scholar
  10. 10.
    Santini C, Brennan D, Mennuni C, Hoess R, Nicosia A, Cortese R, Luzzago A (1998) Efficient display of an HCV cDNA expression library as C-terminal fusion to the capsid protein D of bacteriophage lambda. J Mol Biol 282:125–135CrossRefGoogle Scholar
  11. 11.
    Beghetto E, De Paolis F, Spadoni A, Del Porto P, Buffolano W, Gargano N (2008) Molecular dissection of the human B-cell response against cytomegalovirus infection by lambda-display. J Virol Methods 151:7–14CrossRefGoogle Scholar
  12. 12.
    Beghetto E, Gargano N, Ricci S, Oggioni M, Garufi G, Peppoloni S, Pozzi G, Felici F (2006) Discovery of a novel Streptococcus pneumoniae antigen by screening a whole genome lambda-display library. FEMS Microbiol Lett 262:14–21CrossRefGoogle Scholar
  13. 13.
    De Paolis F, Beghetto E, Spadoni A, Oggioni M, Montagnani F, Felici F, Gargano N (2007) Identification of a human immunodominant B-cell epitope within IgA1 protease of Streptococcus pneumoniae. BMC Microbiol 7:113CrossRefGoogle Scholar
  14. 14.
    Beghetto E, De Paolis F, Montagnani F, Cellesi C, Gargano N (2009) Discovery of new Mycoplasma pneumoniae antigens by use of a whole-genome lambda-display library. Microbes Infect 11:66–73CrossRefGoogle Scholar
  15. 15.
    Montagnani F, De Paolis F, Beghetto E, Gargano N (2010) Use of recombinant chimeric antigens for the serodiagnosis of Mycoplasma pneumoniae infection. Eur J Clin Microbiol Infect Dis 29(11):1377–1386CrossRefGoogle Scholar
  16. 16.
    Papasergi S, Lanza Cariccio V, Pietrocola G, Domina M, D'Aliberti D, Trunfio MG, Signorino G, Peppoloni S, Biondo C, Mancuso G, Midiri A, Rindi S, Teti G, Speziale P, Felici F, Beninati C (2013) Immunogenic properties of Streptococcus agalactiae FbsA fragments. PLoS One 8(9):e75266CrossRefGoogle Scholar
  17. 17.
    Domina M, Lanza Cariccio V, Benfatto S, D'Aliberti D, Venza M, Borgogni E, Castellino F, Biondo C, D'Andrea D, Grassi L, Tramontano A, Teti G, Felici F, Beninati C (2014) Rapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries. PLoS One 9(12):e114159CrossRefGoogle Scholar
  18. 18.
    Domina M, Lanza Cariccio V, Benfatto S, Venza M, Venza I, Borgogni E, Castellino F, Midiri A, Galbo R, Romeo L, Biondo C, Masignani V, Teti G, Felici F, Beninati F (2016) Functional characterization of a monoclonal antibody epitope using a lambda phage display-deep sequencing platform. Sci Rep 6:31458CrossRefGoogle Scholar
  19. 19.
    Cano PG, Gamage LNA, Marciniuk K, Hayes C, Napper S, Hayes S, Griebel PJ (2017) Lambda display phage as a mucosal vaccine delivery vehicle for peptide antigens. Vaccine 35(52):7256–7263CrossRefGoogle Scholar
  20. 20.
    Fedson DS, Anthony J, Scott G (1999) The burden of pneumococcal disease among adults in developed and developing countries: what is known and what is not known. Vaccine 17:S11–S18CrossRefGoogle Scholar
  21. 21.
    Tuomanen EI (2000) Pathogenesis of pneumococcal inflammation: otitis media. Vaccine 19:S38–S40CrossRefGoogle Scholar
  22. 22.
    Bogaert D, De Groot R, Hermans PWM (2004) Streptococcus pneumoniae colonization: the key to pneumococcal disease. Lancet Infect Dis 4:144–154CrossRefGoogle Scholar
  23. 23.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.IRBM Science ParkRomeItaly
  2. 2.Alfasigma S.p.ARomeItaly

Personalised recommendations