Selection of phage-displayed human antibody fragments on Dengue virus particles captured by a monoclonal antibody: application to the four serotypes.
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.
Cabezas
Sheila
S
Department of Virology, PAHO/WHO Collaborating Center for the Study of Dengue and its Vector, Pedro Kourí Tropical Medicine Institute, Autopista Novia del Mediodía, km 6 5, Marianao 13, Habana, Cuba. sheila@cigb.edu.cu
Rojas
Gertrudis
G
Pavon
Alequis
A
Alvarez
Mayling
M
Pupo
Maritza
M
Guillen
Gerardo
G
Guzman
Maria G
MG
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D001435:Bacteriophages; D002460:Cell Line; D003429:Cross Reactions; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D019151:Peptide Library; D012703:Serotyping
D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D019151:Peptide Library