Selection and characterization of scFv antibodies against the Sin Nombre hantavirus nucleocapsid protein.
Rodent-borne hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the old world and hantavirus cardio-pulmonary syndrome (HCPS) in the new. Most cases of HCPS in North America are caused by Sin Nombre Virus (SNV). Current viral detection technologies depend upon the identification of anti-viral antibodies in patient serum. Detection of viral antigen may facilitate earlier detection of the pathogen. We describe here the characterization of two single-chain Fv antibodies (scFvs), selected from a large naïve phage antibody library, which are capable of identifying the Sin Nombre Virus nucleocapsid protein (SNV-N), with no cross reactivity with the nucleocapsid protein from other hantaviruses. The utility of such selected scFvs was increased by the creation of an scFv-alkaline phosphatase fusion protein which was able to directly detect virally produced material without the need for additional reagents.
Velappan
Nileena
N
Los Alamos National Laboratory, TA-43, HRL-1, MS M888, Los Alamos NM 87545, USA.
Martinez
Jennifer S
JS
Valero
Rosa
R
Chasteen
Leslie
L
Ponce
Liana
L
Bondu-Hawkins
Virginie
V
Kelly
Craig
C
Pavlik
Peter
P
Hjelle
Brian
B
Bradbury
Andrew R M
AR
D000469:Alkaline Phosphatase / Q000235:genetics / Q000276:immunology; D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D018804:Hantavirus Pulmonary Syndrome / Q000175:diagnosis / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D029261:Sin Nombre virus / Q000235:genetics / Q000276:immunology*; D014709:Vero Cells
D000914:Antibodies, Viral; D007135:Immunoglobulin Variable Region; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase