摘要

The major concern for severe acute respiratory syndrome (SARS), caused by the SARS-associated coronavirus (SARS-CoV), is the lack of diagnostic and therapeutic agents. Using a phage display technology in a chicken system, high-affinity monoclonal antibody fragments against the SARS-CoV spike protein were characterized. Ten truncated spike protein gene fragments were expressed in Escherichia coli cells. Following the immunization of chickens with these recombinant spike proteins, two single-chain variable fragment (scFv) antibody libraries were established with short or long linkers to contain 5 × 10 7 and 9 × 10 6 transformants, respectively. After four rounds of panning selection, the scFv antibodies of randomly chosen clones were demonstrated by Coomassie blue staining, and verified by western blot analysis. In a comparison of nucleotide sequences with the chicken germline gene, we found that all clones varied in the complementarity-determining regions, that two scFv antibodies reacted significantly with SARS-CoV-infected Vero cells, and that those two specific scFv antibodies recognized the same region of the spike protein spanning amino acid residues 750-1000. In conclusion, the results suggest that the chicken scFv phage display system can be a potential model for mass production of high-affinity antibodies against the SARS-CoV spike protein.
原文英語
頁(從 - 到)104-111
頁數8
期刊Journal of Virological Methods
146
發行號1-2
DOIs
出版狀態已發佈 - 十二月 2007

指紋

Severe Acute Respiratory Syndrome
Single-Chain Antibodies
Chickens
Proteins
Bacteriophages
Clone Cells
Complementarity Determining Regions
Immunoglobulin Fragments
Coronavirus
Vero Cells
Antibody Affinity
Recombinant Proteins
Immunization
Western Blotting
Monoclonal Antibodies
Staining and Labeling
Escherichia coli
Technology
Amino Acids
Genes

ASJC Scopus subject areas

  • Virology

引用此文

Chicken single-chain variable fragments against the SARS-CoV spike protein. / Lee, Yu Ching; Leu, Sy Jye Christine; Hu, Chaur Jong; Shih, Neng Yao; Huang, I. Jen; Wu, Hsueh Hsia; Hsieh, Wen Shyang; Chiang, Bor Luen; Chiu, Wen Ta; Yang, Yi Yuan.

於: Journal of Virological Methods, 卷 146, 編號 1-2, 12.2007, p. 104-111.

研究成果: 雜誌貢獻文章

Lee, Yu Ching ; Leu, Sy Jye Christine ; Hu, Chaur Jong ; Shih, Neng Yao ; Huang, I. Jen ; Wu, Hsueh Hsia ; Hsieh, Wen Shyang ; Chiang, Bor Luen ; Chiu, Wen Ta ; Yang, Yi Yuan. / Chicken single-chain variable fragments against the SARS-CoV spike protein. 於: Journal of Virological Methods. 2007 ; 卷 146, 編號 1-2. 頁 104-111.
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