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.
Liu
Hui
H
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
Ding
Yan-Li
YL
Han
Wei
W
Liu
Mei-Yun
MY
Tian
Rui-Yang
RY
Yang
Sheng-Li
SL
Gong
Yi
Y
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