A recombinant Fab neutralizes dengue virus in vitro.
A recombinant Fab that recognizes a neutralizing epitope located in the (296-400) region of protein E of dengue virus was obtained from cloned hybridoma cells secreting the mouse monoclonal antibody (mAb) 4E11. The Fd and light chain antibody genes were amplified by polymerase chain reaction, cloned into the phagemid vector pMad, expressed in bacteria to produce Fab fragments and sequenced. The mAb 4E11, in particular its light chain complementary-determining regions, shared homologies with two other anti-viral mAbs. The affinity of the parental mAb and the cloned Fab to the MalE-E(296-400) fusion protein were shown to be of the same magnitude, i.e. nanomolar. Fab 4E11 neutralization capacity was found between 8 and 4-times or less lower than that of mAb 4E11, depending on serotypes, thus the Fab could have a smaller antiviral activity than the mAb in vitro.
Thullier
P
P
Laboratoire d'Ingéniérie des Anticorps, Institut Pasteur, Paris, France.
Lafaye
P
P
Mégret
F
F
Deubel
V
V
Jouan
A
A
Mazié
J C
JC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000276:immunology; D003001:Cloning, Molecular; D003716:Dengue Virus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006825:Hybridomas; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D014759:Viral Envelope Proteins / Q000276:immunology*; D010948:Viral Plaque Assay
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C072281:E protein TH Sman, Dengue virus