Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV

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• 作者：Aizhi Zhao ; Weijun Qin ; Yueheng Han ; Weihong Wen ; Wenhong Zhang ; Zhonghui Lian ; Gang Chen ; Zhuoli Zhang ; Jianqiang Peng ; He Wang ; et al.
• 关键词：SARS-CoV ; scFv ; Nucleocapsid ; Phage display
• 刊名：Microbes and Infection
• 出版年：2007
• 出版时间：July 2007
• 年：2007
• 卷：9
• 期：8
• 页码：1026-1033
• 全文大小：248 K

文摘

To develop reagents for early diagnosis and therapeutic drugs against SARS-associated coronavirus (SARS-CoV), a large (3 × 10⁹) immunized human antibody library was constructed from peripheral blood mononuclear cells from six SARS convalescent patients. A single chain variable fragment antibody (N18) with high affinity against N protein of SARS-CoV was isolated. Sequence analysis revealed that the VL gene was composed of VL3 h (V lambda subgroup) and JL2 regions and the VH gene was composed of VH1-69 (VH1 subgroup), D2-15, D3-22 and JH6 regions. Soluble N18 antibody was expressed in Escherichia coli HB2151, purified by Ni–NTA affinity chromatography and verified by SDS-PAGE and Western blot. The potential application for early diagnosis was evaluated using N protein capture ELISA in which N18 antibody demonstrated high sensitive activity in detecting N protein of SARS-CoV. Finally, the potential usefulness of the N18 antibody in prophylaxis, vaccine design and therapy of SARS is discussed.