Recombinant scFv antibodies against E protein and N protein of severe acute respiratory syndrome virus.
Three single chain antibodies (scFv) against the proteins of severe acute respiratory syndrome coronavirus (SARS-CoV) were isolated by phage display from an scFv antibody library. Bio-panning was carried out against immobilized purified envelope (E) and nucleocapsid (N) proteins of SARS-CoV. Their binding activity and specificity to E or N protein of SARS-CoV were characterized by phage-ELISA. Two of them, B10 and C20, could recognize non-overlapping epitopes of the E protein according to the two-site binding test result. Clone A17 could recognize N protein. The sequence of the epitope or overlapping epitope of scFv antibody A17 was PTDSTDNNQNGGRNGARPKQRRPQ. The affinity (equilibrium dissociation constant, K(d)) of SARS-CoV E protein was 5.7 x 10(-8) M for B10 and 8.9 x 10(-8) M for C20. The affinity of A17 for N protein was 2.1 x 10(-6) M. All three scFv antibodies were purified with affinity chromatography and determined by Western blot.
Shanghai Research Center of Biotechnology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200233, China. huiliu@srcb.ac.cn
Hui
H
Liu
Yan-Li
YL
Ding
Wei
W
Han
Mei-Yun
MY
Liu
Rui-Yang
RY
Tian
Sheng-Li
SL
Yang
Yi
Y
Gong
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000378:metabolism; D000915:Antibody Affinity; D000086462:Coronavirus Nucleocapsid Proteins; D015686:Gene Products, env / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000276:immunology*; D019151:Peptide Library; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*
D000914:Antibodies, Viral; D000086462:Coronavirus Nucleocapsid Proteins; D015686:Gene Products, env; D007128:Immunoglobulin Fragments; D019590:Nucleocapsid Proteins; D019151:Peptide Library; C055966:immunoglobulin Fv