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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 5.7´108 M for B10 and 8.9´108 M for C20. The affinity of A17 for N protein was 2.1´106 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]