[Transient expression and characterization of intracellular single chain Fv against the nucleocapsid protein of Hantavirus].
OBJECTIVE
To transiently express an intracellular single chain Fv of monoclonal antibody 1A8 against nucleocapsid protein of Hantavirus and characterize the immunological activities of the expressed products.


METHODS
COS-7 cells were transfected with mammalian expression vector 1A8-scFv-Ckappa/pCI-neo via lipofectin. The expressed product was identified by indirect immunofluorescence and immunoprecipitation.


RESULTS
A diffuse pattern fluorescence was observed in less than 1% cytoplasm of transfected COS-7 cells. The binding of intracellular antibody fragments to NP antigen was confirmed by immunoprecipitation analysis.


CONCLUSIONS
Transiently expressed single chain intrabodies can effectively target NP antigen in the cytoplasm. The present study may provide a new approach for treatment of Hantavirus.
Bai
Wen-tao
WT
Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
Xu
Zhi-kai
ZK
Zhang
Fang-lin
FL
Luo
Wen
W
Liu
Yong
Y
Wu
Xing-an
XA
Yan
Yan
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D019556:COS Cells; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D005822:Genetic Vectors; D006476:Orthohantavirus / Q000235:genetics / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000276:immunology / Q000378:metabolism*; D014162:Transfection
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D019590:Nucleocapsid Proteins