國家衛生研究院 NHRI:Item 3990099045/2698
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    题名: Chicken single-chain variable fragments against the SARS-CoV spike protein
    作者: Lee, YC;Leu, SJC;Hu, CJ;Shih, NY;Huang, IJ;Wu, HH;Hsieh, WS;Chiang, BL;Chiu, WT;Yang, YY
    贡献者: National Institute of Cancer Research
    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 x 10(7) and 9 x 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. (c) 2007 Elsevier B.V. All rights reserved.
    关键词: Biochemical Research Methods;Biotechnology & Applied Microbiology;Virology
    日期: 2007-12
    關聯: Journal of Virological Methods. 2007 Dec;146(1-2):104-111.
    Link to: http://dx.doi.org/10.1016/j.jviromet.2007.06.010
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0166-0934&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000251648400014
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=35648935867
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