Taipei Medical University Institutional Repository:Item 987654321/3736
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    题名: Chicken single-chain variable fragments against the SARS-CoV spike protein
    作者: 吳雪霞;楊沂淵
    Lee Yu-Ching;Leu Sy-Jye C;Hu Chaur-Jong;Shih Neng-Yao;Huang I-Jen;Wu Hsueh-Hsia;Hsieh Wen-Shyang;Chiang Bor-Luen
    贡献者: 醫學檢驗暨生物技術學系
    关键词: SARS-CoV;Phage display technology;Spike protein;scFv
    日期: 2007
    上传时间: 2009-08-25 10:37:59 (UTC+8)
    摘要: Abstract
    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 5×107 and 9×106 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.
    關聯: Journal of Virological Methods.(146):104-111.
    数据类型: article
    显示于类别:[醫學檢驗暨生物技術學系所] 期刊論文

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