Neutralizing human single-chain antibodies against Herpes Simplex Virus type 1 glycoprotein D from a phage display library
نویسندگان
چکیده
Among the 12 glycoproteins of the Herpes simplex virus type 1 envelope (HSV-1), glycoprotein D (gD) plays a critical role in the entry of the virus into target cells and cell-to-cell spread. gD is an attractive target for molecular intervention and monoclonal antibodies to this glycoprotein has decreased the severity of experimental HSV-1 infection in animal studies. Single chain antibodies which are produced by antibody engineering are small high affinity recombinant antibodies with growing clinical importance especially when viral antigens have been used for targeted therapy. Here we report neutralizing singlechain fragment variable (scFv) antibodies against HSV-1 gD from a phage-display non-immune human scFv library. The phage antibody was panned against amino acid residues 12-21 derived from the Nterminal part of gD. Two scFvs, scFv-gD1 and scFv-gD2, with frequencies of 25% and 20% were isolated among scFv clones using PCR and MvaI fingerprinting. Phage ELISA analysis demonstrated high reactivity of scFv-gD1 and scFv-gD2 with the gD peptide. In neutralization assay, scFv-gD2 exhibited neutralizing effect of 76%. Sequence analysis of scFv-gD2 revealed the amino acid specific changes in FR1 region of heavy chain and FR1 and JL3 regions of light chain of antibody molecule. Also the sequence GADTAMAG in CDR3 region represented the specificity of the selected antibody. The results suggest that the specific neutralizing scFv-gD2 can be considered as a new alternative in the prophylaxis and treatment of HSV-1 infections.
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