Development of a monoclonal antibody specific to a recombinant envelope protein from dengue virus type 4 expressed in Pichia pastoris.
A mouse monoclonal antibody (MAb, 4B6) was able to recognize dengue virus type 4 envelope (E) protein both as a recombinant protein in Pichia pastoris and when it was present in infected brains of suckling mice. 4B6 was characterized by enzyme-linked immunoadsorbent assay (ELISA), hemaglutination inhibition, neutralization, and immunoblot. The MAb was isotyped as IgG2a. It was serotype 4 specific and it inhibited hemaglutination and neutralized homologous virus. It did not enhance infection of P338D1 cells by dengue type 4 virus strain H-241 strain. This MAb was reactive with recombinant E protein and dengue 4 virus, as revealed by Western blot. In vivo, MAb 4B6 conferred passive protection in mice challenged with homologous virus. Currently, this MAb is being used to purify recombinant E protein for further studies.
Pupo-Antúnez
M
M
Virology Department, Tropical Medicine Institute, Pedro Kouri, Havana City, Cuba. mpupo@ipk.sld.cu
Rodriguez
R
R
Alvarez
M
M
Amin
N
N
Rodriguez
H
H
Otero
A
A
Guzmán
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000627:therapeutic use; D000914:Antibodies, Viral / Q000276:immunology* / Q000627:therapeutic use; D000918:Antibody Specificity; D003715:Dengue / Q000628:therapy*; D003716:Dengue Virus / Q000276:immunology*; D005260:Female; D006825:Hybridomas; D007116:Immunization, Passive; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins