Selection of single-chain antibodies that specifically interact with vesicular stomatitis virus (VSV) nucleocapsid and inhibit viral RNA synthesis.
The RNA genome of non-segmented negative-strand RNA viruses is completely covered by the nucleoprotein (N) forming a ribonucleoprotein complex, the nucleocapsid. The nucleocapsid functions as the template for viral RNA synthesis that is mediated by a viral RNA-dependent RNA polymerase. It is postulated that the selection of molecules that would specifically target the nucleocapsid and thus inhibit the viral polymerase activity could represent a common approach to block negative-strand RNA viruses. Two single-chain antibody fragments (scFv) that were selected using the phage display technology and interacted specifically with vesicular stomatitis virus (VSV) nucleocapsid were characterized. The two recombinant antibodies recognize a conformational epitope on the nucleocapsid and immunoprecipitate specifically nucleocapsids from infected cell extracts. Both antibodies have a strong inhibitory effect on VSV transcription activity in vitro. Thus, they represent starting molecules for future development of in vivo viral RNA synthesis inhibitors.
Cortay
Jean-Claude
JC
Immunité & Infections Virales, CNRS, Université Lyon 1 UMR 5537, IFR Laennec, 69372 Lyon Cedex 08, France.
Gerlier
Denis
D
Iseni
Frédéric
F
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology*; D000918:Antibody Specificity; D000939:Epitopes / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000276:immunology* / Q000494:pharmacology*; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000235:genetics / Q000276:immunology*; D019151:Peptide Library; D012367:RNA, Viral / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology; D014158:Transcription, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000276:immunology* / Q000378:metabolism
D000914:Antibodies, Viral; D000939:Epitopes; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D012367:RNA, Viral; D011994:Recombinant Proteins