Mapping of antigenic determinants on a SAT2 foot-and-mouth disease virus using chicken single-chain antibody fragments.
Recombinant single-chain variable fragments (scFvs) of antibodies make it possible to localize antigenic and immunogenic determinants, identify protective epitopes and can be exploited for the design of improved diagnostic tests and vaccines. A neutralizing epitope, as well as other potential antigenic sites of a SAT2 foot-and-mouth disease virus (FMDV) were identified using phage-displayed scFvs. Three unique ZIM/7/83-specific scFvs, designated scFv1, scFv2 and scFv3, were isolated. Further characterization of these scFvs revealed that only scFv2 was capable of neutralizing the ZIM/7/83 virus and was used to generate neutralization-resistant virus variants. Sequence analysis of the P1 region of virus escaping neutralization revealed a residue change from His to Arg at position 159 of the VP1 protein. Residue 159 is not only surface exposed but is also located at the C-terminal base of the G-H loop, a known immunogenic region of FMDV. A synthetic peptide, of which the sequence corresponded to the predicted antigenic site of the VP1 G-H loop of ZIM/7/83, inhibited binding of scFv2 to ZIM/7/83 in a concentration-dependent manner. This region can therefore be considered in the design of SAT2 vaccine seed viruses for the regional control of FMD in Africa.
Onderstepoort Veterinary Institute, Transboundary Animal Diseases Programme, Private Bag X05, Onderstepoort 0110, South Africa. StoreyP@arc.agric.za
Pamela A
PA
Opperman
Francois F
FF
Maree
Wouter
W
Van Wyngaardt
Wilna
W
Vosloo
Jacques
J
Theron
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D002645:Chickens; D006224:Cricetinae; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000276:immunology*
D000914:Antibodies, Viral; D000939:Epitopes; D057127:Single-Chain Antibodies