Human Single-Chain Fv Antibodies Against Burkholderia mallei and Burkholderia pseudomallei

Abstract

Much effort has been devoted to the development of mouse monoclonal antibodies that react specifically with Burkholderia mallei and Burkholderia pseudomallei for diagnostic and/or therapeutic purposes. Our present study focused on the screening of a phage-displayed nonimmune human single-chain Fv (scFv) antibody library against heat-killed B. mallei and B. pseudomallei for the generation of human scFv antibodies specific to the two pathogenic species of bacteria. Using two different panning procedures, we obtained seven different scFv phage antibodies that interacted with the heat-killed whole bacterial cells of B. mallei and B. pseudomallei. Our results demonstrate that panning of a human scFv antibody library against heat-killed whole bacterial cells may provide a valuable strategy for developing human monoclonal antibodies against the highly pathogenic bacteria.

Get full access to this article

View all access options for this article.

References

Anuntagool N, Sirisinha S. Antigenic relatedness between Burkholderia pseudomallei and Burkholderia mallei. Microbiol Immunol 46:143–150, 2002.

Leelarasamee A. Recent development in melioidosis. Curr Opin Infect Dis 17:131–136, 2004.

Woods DE, DeShazer D, Moore RA, Brett PJ, Burtnick MN, Reckseidler SL, Senkiw MD. Current studies on the pathogenesis of melioidosis. Microbes Infect 1:157–162, 1999.

Jeddeloh LA, Fritz DL, Waag DM, Hartings JM, Andrews GP. Biodefense-driven murine model of pneumonic melioidosis. Infect Immun 71:584–587, 2003.

Jones SM, Ellis JF, Russell P, Griffin KF, Oyston PC. Passive protection against Burkholderia pseudomallei infection in mice by monoclonal antibodies against capsular polysaccharide, lipopolysaccharide or proteins. J Med Microbiol 51:1055–1062, 2002.

Ho M, Schollaardt T, Smith MD, Perry MB, Brett PJ, Chaowagul W, Bryan LE. Specificity and functional activity of anti-Burkholderia pseudomallei polysaccharide antibodies. Infect Immun 65:3648–3653, 1997.

Pongsunk S, Thirawattanasuk N, Piyasangthong N, Ekpo P. Rapid identification of Burkholderia pseudomallei in blood cultures by a monoclonal antibody assay. J Clin Microbiol 37:3662–3667, 1999.

Steinmetz I, Reganzerowski A, Brenneke B, Haussler S, Simpson A, White NJ. Rapid identification of Burkholderia pseudomallei by latex agglutination based on an exopolysaccharide-specific monoclonal antibody. J Clin Microbiol 37:225–228, 1999.

Hoogenboom HR. Overview of antibody phage-display technology and its application. In: O’Brien PM, Aitken R, Eds. Antibody Phage Display: Methods and protocols. Totowa, NJ: Humana Press, pp. 1–37, 2002.

10.

Rader C, Barbas CF 3rd. Phage display of combinatorial antibody libraries. Curr Opin Biotechnol 8:503–508, 1997.

11.

McCafferty J, Griffiths AD, Winter G, Chiswell DJ. Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348: 552–554, 1990.

12.

Lennard S. Standard protocols for the construction of scFv libraries. In: O’Brien PM, Aitken R, Eds. Antibody Phage Display: Methods and protocols. Totowa, NJ: Humana Press, pp. 59–71, 2002.

13.

Nathan S, Li H, Mohamed R, Embi N. Phage display of recombinant antibodies toward Burkholderia pseudomallei exotoxin. J Biochem Mol Biol Biophys 6:45–53, 2002.

14.

Gao C, Mao S, Kaufmann G, Wirsching P, Lerner RA, Janda KD. A method for the generation of combinatorial antibody libraries using pIX phage display. Proc Natl Acad Sci U S A 99:12612–12616, 2002.

15.

Paoli GC, Chen CY, Brewster JD. Single-chain Fv antibody with specificity for Listeria monocytogenes. J Immunol Methods 289:147–155, 2004.

16.

Reiche N, Jung A, Brabletz T, Vater T, Kirchner T, Faller G. Generation and characterization of human monoclonal scFv antibodies against Helicobacter pylori antigens. Infect Immun 70:4158–4164, 2002.

17.

Stacy JE, Kausmally L, Simonsen B, Nordgard SH, Alsoe L, Michaelsen TE, Brekke OH. Direct isolation of recombinant human antibodies against group B Neisseria meningitidis from scFv expression libraries. J Immunol Methods 283:247–259, 2003.

18.

Zhou B, Wirsching P, Janda KD. Human antibodies against spores of the genus Bacillus: a model study for detection of and protection against anthrax and the bioterrorist threat. Proc Natl Acad Sci U S A 99: 5241–5246, 2002.