Phage-displayed and soluble mouse scFv fragments neutralize rabies virus.
A phage-display technology was used to produce a single-chain Fv antibody fragment (scFv) from the 30AA5 hybridoma secreting anti-glycoprotein monoclonal antibody (MAb) that neutralizes rabies virus. ScFv was constructed and then cloned for expression as a protein fusion with the g3p minor coat protein of filamentous phage. The display of antibody fragment on the phage surface allows its selection by affinity using an enzyme-linked immunosorbent assay (ELISA); the selected scFv fragment was produced in a soluble form secreted by E. coli. The DNA fragment was sequenced to define the germline gene family and the amino-acid subgroups of the heavy (VH) and light (VL) chain variable regions. The specificity characteristics and neutralization capacity of phage-displayed and soluble scFv fragments were found to be identical to those of the parental 30AA5 MAb directed against antigenic site II of rabies glycoprotein. Phage-display technology allows the production of new antibody molecule forms able to neutralize the rabies virus specifically. The next step could be to engineer and produce multivalent and multispecific neutralizing antibody fragments. A cocktail of multispecific neutralizing antibodies could contain monovalent, bivalent or tetravalent scFv fragments, for passive immunoglobulin therapy.
Muller
B H
BH
Laboratoire de Prédéveloppement des Sondes, Institut Pasteur, Paris, France.
Lafay
F
F
Demangel
C
C
Perrin
P
P
Tordo
N
N
Flamand
A
A
Lafaye
P
P
Guesdon
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D001666:Binding Sites, Antibody; D003001:Cloning, Molecular; D006224:Cricetinae; D000939:Epitopes / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D017911:Inoviridae / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011820:Rabies virus / Q000276:immunology*; D017422:Sequence Analysis, DNA; D012995:Solubility
D000939:Epitopes; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region