Selection of phage-displayed human antibody fragments on Dengue virus particles captured by a monoclonal antibody: application to the four serotypes

J Virol Methods. 2008 Feb;147(2):235-43. doi: 10.1016/j.jviromet.2007.09.001. Epub 2007 Oct 22.

Abstract

Antibody fragments to the four Dengue virus serotypes were isolated from a human universal naïve library using phage display technology. Phage-displayed antibody fragments were selected on Dengue virus particles directly captured from infected Vero cells supernatant by an anti-dengue monoclonal antibody, in order to avoid laborious virus concentration/purification procedures. A total of nine phage-displayed antibody fragments were obtained. Seven of them were highly specific for three of the selector serotypes (two for Dengue 1, four for Dengue 3 and one for Dengue 4). One clone (Dengue 3-selected) cross-reacted with Dengue 1, whereas another (selected with Dengue 2) cross-reacted with the three remaining serotypes. The soluble variants of six antibody fragments recognized their target viruses when used at nanomolar and even subnanomolar concentrations. All phage-displayed antibody fragments were cross-reactive against several strains of distinct genotypes within the corresponding serotype(s). These antibody fragments are potentially useful for the future development of tools for viral diagnosis and serotype identification. The simple phage selection method on captured virus could be applied in a high throughput way to obtain larger panels of antibody fragments to Dengue virus for multiple applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / genetics
  • Antibodies, Viral / immunology*
  • Antibody Specificity
  • Bacteriophages
  • Cell Line
  • Cross Reactions
  • Dengue Virus / classification
  • Dengue Virus / immunology*
  • Humans
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / immunology*
  • Peptide Library*
  • Serotyping

Substances

  • Antibodies, Viral
  • Immunoglobulin Fragments
  • Peptide Library