Abstract
Polyclonal antibodies, as well as monoclonal antibodies are efficacious in providing protective immunity against Francisella tularensis. This study demonstrates the application of phage display libraries for the construction of monoclonal antibodies against F. tularensis. Novel single-chain fragment variable (scFv) antibodies were generated against a whole bacterial lysate of F. tularensis live vaccine strain using the human single fold scFv libraries I (Tomlinson I + J). A total of 20 clones reacted with the bacterial cell lysate. Further, the library contains two clones responsive to recombinant lipoprotein FTT1103Δsignal (F. tularensis subsp. tularensis Schu S4), which was constructed without a signal sequence. These positively-binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA). Then, positive scFvs were expressed in a soluble form in Escherichia coli HB2151 and tested for positive scFvs by using scFv-ELISA.
Article PDF
Avoid common mistakes on your manuscript.
References
Köhler G., Milstein C., Continuous cultures of fused cells secreting antibody of predefined specificity, Nature, 1975, 256, 495–497
Winter G., Milstein C., Man-made antibodies, Nature, 1991, 349, 293–299
Lonberg N., Human antibodies from transgenic animals, Nat. Biotechnol., 2005, 23, 1117–1125
Smith G.P., Filamentous fusion phage, novel expression vectors that display cloned antigens on the virion surface, Science, 1985, 228, 1315–1317
Barbas C.F. 3rd, Kang A.S., Lerner R.A., Benkovic S.J., Assembly of combinatorial antibody libraries on phage surfaces, the gene III site, Proc. Natl. Acad. Sci. USA, 1991, 88, 7978–7982
Hoogenboom H.R., Overview of antibody phage-display technology and its applications, Meth. Mol. Biol., 2002, 178, 1–37
Scott J.K., Smith G.P., Searching for peptide ligands with an epitope library, Science, 1990, 249, 386–390
Szardenings M., Phage display of random peptide libraries: applications, limits, and potential, J. Recept. Signal Transduct. Res., 2003, 23, 307–349
Clackson T., Hoogenboom H.R., Griffiths A.D., Winter G., Making antibody fragments using phage display libraries, Nature, 1991, 352, 624–628
Parmley S.F., Smith G.P., Filamentous fusion phage cloning vectors for the study of epitopes and design of vaccines, Adv. Exp. Med. Biol., 1989, 251, 215–218
Vaughan T.P., Williams A.W., Pritchard K., Osbourn J.K., Pope A.R., McCafferty J., et al., Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library, Nat. Biotechnol., 1996, 14, 309–314
Sheets M.D., Amersdorfer P., Finnern R., Sargent P., Lindqvist E., Schier R., et al., Efficient construction of a large nonimmune phage antibody library, the production of high-affinity human single-chain antibodies to protein antigens, Proc. Natl. Acad. Sci. USA, 1998, 95, 6157–6162
Ward E.S., Gussow D., Griffiths A.D., Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli, Nature, 1989, 341, 484–485
Vaughan T.J., Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library, Nat. Biotechnol., 1996, 14, 309–314
Cai X.H., Garen A., Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells: selection of pecific antibodies from single-chain Fv fusion phage libraries, Proc. Natl. Acad. Sci. USA, 1995, 92, 6537–6541
Griffith A.D., Williams S.C., Hartley O., Tomlinson I.M., Waterhouse P., Crosby W.L., et al., Isolation of high affinity human antibodies directly from large synthetic repertoires, EMBO J., 1994, 13, 3245–3260
Greenwood J., Willis A.E., Perham R.N., Multiple display of foreign peptides on a filamentous bacteriophage, Peptides from Plasmodium falciparum circumsporozoite protein as antigens, J. Mol. Biol., 1991, 220, 821–827
Conlan J.W., North R.J., Early pathogenesis of infection in the liver with the facultative intracellular bacteria Listeria monocytogenes, Francisella tularensis, and Salmonella typhimurium involves lysis of infected hepatocytes by leukocytes, Infect. Immun., 1992, 60, 5164–5171
Thorpe B.D., Marcus S., Comparison of two techniques to study in vitro uptake and fate of Pasteurella tularensis, J. Reticuloendothel. Soc., 1964, 15, 418–422
Clemens D.L, Horwitz M.A., Uptake and intracellular fate of Francisella tularensis in human macrophages, Ann. NY Acad. Sci., 2007, 1105, 160–186
Burke D.S., Immunization against tularemia, analysis of the effectiveness of live Francisella tularensis vaccine in prevention of laboratory-acquired tularemia, J. Infect. Dis., 1977, 135, 55–60
Antony L.S.D., Kongshavn P.A.L., Experimental murine tularemia caused by Francisella tularensis, live vaccine strain, a model of acquired cellular resistence, Microb. Pathog., 1987, 2, 3–14
Eigelsbach H.T., Downs C.M., Prophylactic effectiveness of live and killed tularemia vaccines, I. Production of vaccine and evaluation in the white mouse and guinea pig, J. Immunol., 1961, 87, 415–425
Saslaw S., Eigelsbach H.T., Prior J., Wilson H., Carhart S., Tularemia vaccine study II., Intracutaneous study, Arch. Intern. Med., 1961, 107, 121–133
Saslaw S., Eigelsbach H.T., Prior J., Wilson H., Carhart S., Tularemia vaccine study II, Respiratory challenge, Arch. Intern. Med., 1961, 107, 702–714
Qin A., Scott D.W., Thompson J.A., Mann B.J., Identification of an essentials Francisella tularensis subsp. tularensis virulence factor, Infect. Immun., 2009, 77, 152–161
Janovska S., Pavkova I., Reichelova M., Hubalek M., Stulik J., Macela A., Proteomic analysis of antibody response in a case of laboratory-acquired infection with Francisella tularensis subsp. tularensis, Folia Microbiol., 2007, 52, 194–198
Ellermeier C.D., Slauch J.M., RtsA coordinately regulates DsbA and the Salmonella pathogenicity island 1 type III secretion system, J. Bacteriol., 2004, 186, 68–79
Ha U.H., Wang Y., Jin S., DsbA of Pseudomonas aeruginosa is essentials for multiple virulence factors, Infect. Immun., 2003, 71, 1590–1595
Hayashi S., Abe M., Kimoto M., Furukawa S., Nakazawa T., The dsbA-dsbB disulfide bond formation system of Burkholderia cepacia is involved in the production of protease and alkaline phosphatase, motility, metal resistence, and multidrug resistence, Microbiol. Immunol., 2000, 44, 41–50
Miki T., Okada N., Danbara H., Two periplasmatic disufide oxidoreductases, DsbA and SrgA, target outer membrane protein SpiA, a component of the Samonella pathogenicity island 2 type III secretion system, J. Biol. Chem., 2004, 279, 34631–34642
Takatsuka Y., Nikaido H., Site-directed disulfide cross-linking show that cleft flexibility in the periplasmic domain is needed for the multidrug efflux pump AcrB of Escherichia coli, J. Bacteriol., 2007, 189, 8577–8584
Yu J., Inactivation of DsbA, but not DsbC and DsbD, affects the intracellular survival and virulence of Shigella flexneri, Infect. Immun., 1998, 66, 3909–3917
Thakran S., Li H., Lavine C.L., Miller M.A., Bina J.E., Bina X.R., et al., Identification of Francisella tularensis lipoproteins that stimulate the toll-like receptor (TLR)2/TLR1 heterodimer, J. Biol. Chem., 2008, 283, 3751–3760
Sambrook J., Russel D.W., Molecular Cloning — a laboratory manual, 3rd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2001
Zacher A.N. 3rd, Stock C.A., Golden J.W. 2nd, Smith G.P., A new filamentous phage cloning vector, fd-tet, Gene, 1980, 1–2, 127–140
Inaba K., Ito K., Structure and mechanism of the DsbB-DsbA disulfide bond generation machine, Biochim. Biophys. Acta, 2008, 1783, 520–529
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Kubelkova, K., Macela, A. Development of tularemic scFv antibody fragments using phage display. cent.eur.j.biol. 5, 310–317 (2010). https://doi.org/10.2478/s11535-010-0015-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11535-010-0015-3