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    Please use this identifier to cite or link to this item: http://ir.cnu.edu.tw/handle/310902800/21334


    標題: 霍亂毒素多株抗體之製備與純化
    Production and Purification of Polyclonal Antibodies Against Cholera Toxin
    作者: 周淑芬
    邱慶豐
    王淑珍
    張竣凱
    貢獻者: 生物科技系
    食品科技系
    關鍵字: 霍亂毒素
    多株抗體
    酵素連結免疫分析法
    Cholera toxin
    Polyclonal antibodies
    Enzyme-linked immunosorbent assay(ELISA)
    v30_101_108.pdf
    日期: 2004
    上傳時間: 2009-06-01 11:41:37 (UTC+8)
    摘要: 本研究目的在使用霍亂毒素(cholera toxin, CT)腹腔免疫ICR小鼠,來生產霍亂毒素之多株抗體(cholera toxin polyclonal antibodies, anti-CT)。將高度免疫(hyperimmune)之ICR小鼠,注射0.5 ml pristane後,再間隔兩週注射106個NS-1骨髓癌細胞。收集ICR小鼠所產生之腹水及血清,利用Hitrap rProtein A column 來純化多株抗體,並進一步分析已純化anti-CT之抗體效價及其專一性。本研究以間接型酵素連結免疫分析法(enzymelinkedimmunosorbent assay, indirect ELISA)測定anti-CT之效價。最後進行anti-CT交叉反應之測定,採用志賀菌Shigella dysenteriae 1(S.D., ATCC 13313),腸道出血型大腸桿菌(enterohemorrhagic E. coli, EHEC, E. coli O157:H7, BCRC 13086)以及腸道毒素型大腸桿菌(enterotoxigenic E. coli, ETEC, WHO 110)所分泌之腸內毒素進行測試。實驗結果顯示,多株抗體anti-CT的效價為1:1,000。可測得霍亂毒素最低濃度在0.01 µg/ml ( CT濃度在0.01~100 µg/ml與吸光值呈線性關係)。當所培養之三種腸內桿菌菌數達106CFU/ml時,取其上層液進行測試,結果發現與本研究所生產之anti-CT均有低交叉反應。
    The aim of this program was to produce polyclonal antibodies against cholera toxin (anti-CT), from Vibrio cholerae. ICR mice was immunized intraperitoneally with cholera toxin (CT) for four to five times every four weeks. Hyperimmune ICR mice produced anti-CT after injecting with 0.5 ml pristane, and injected with NS-1 myeloma cells two weeks later. Ascites and serum were collected from ICR mice, and anti-CT was purified by using Hitrap rProtein A column, and the titer and specificity of anti-CT were determined by indirect enzyme-linked immunosorbent assay (ELISA). In cross-reaction experiment for anti-CT with other enterotoxins, containing enterotoxins of Shigella dysenteriae 1 (S.D., ATCC 13313), enterohemorrhagic E. coli (EHEC, CCRC 13086), and enterotoxigenic E. coli (ETEC, WHO 110) was measured. The result showed the titer of anti-CT was 1:1,000, and had low cross-reaction with three kinds of enterotoxins from 106 CFU/ml enterobacteria.
    Appears in Collections:[嘉南學報] 30 期 (2004)
    [生物科技系(所)] 期刊論文
    [ 食品科技系 ] 期刊論文

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