Generation and characterization of human monoclonal antibodies to G5, a linear neutralization epitope on glycoprotein of rabies virus, by phage display technology.
The aim of the present study was to discover distinct human MAbs to RV with high neutralizing potency and a broad neutralization spectrum. A phage display technology was used to produce human scFv to G5, a conserved linear neutralization epitope on Gp of RV. A phage display scFv library with 6 x 10(7) members was constructed and the phage-scFv with 'antigen-binding' activities were selected with synthetic peptide G5-24. The obtained scFv genes were cloned into pET22b(+)/BL21(DE3) and from this we prepared purified scFv fragments. The assay of the specificity characteristics and neutralization capacity showed that two distinct clones with new human immunoglobulin V genes can recognize G5 specifically as well as neutralize different RV strains. They have potential for inclusion in an antibodies combination aimed for use in rabies PEP.
Zhao
Xiao-Ling
XL
Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Tianjin, China. xlzhao985@sohu.com
Yin
Jun
J
Chen
Wei-Qiang
WQ
Jiang
Ming
M
Yang
Guan
G
Yang
Zhi-Hua
ZH
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D000956:Antigens, Viral / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D011820:Rabies virus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*; D010948:Viral Plaque Assay
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus