https://www.abbs.info E-mail: [email protected]
ISSN
1672-9145
Acta Biochim Biophys Sin
2004, 36(8): 541–547
CN 31-1940/Q
Recombinant scFv Antibodies against E Protein and N Protein of
Severe Acute Respiratory Syndrome Virus
Hui LIU*, Yan-Li DING, Wei
HAN, Mei-Yun LIU, Rui-Yang TIAN, Sheng-Li YANG, and Yi GONG*
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
Abstract 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, Kd) of SARS-CoV E protein was 5.7108 M for B10 and 8.9108 M for C20. The affinity of A17
for N protein was 2.1106 M. All three scFv antibodies
were purified with affinity chromatography and determined by Western blot.
Key words SARS-CoV; antibody; scFv; phage display; affinity chromatography
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Received: April 21, 2004 Accepted: June 23, 2004
*Corresponding authors:
Yi GONG: Tel, 86-21-64700892 ext. 371; Fax, 86-21-64700244; E-mail, [email protected]
Hui LIU: Tel, 86-21-64700892 ext. 371; Fax, 86-21-64700244; E-mail, [email protected]