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題 名 | 霍亂毒素多株抗體之製備與純化=Production and Purification of Polyclonal Antibodies Against Cholera Toxin |
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作 者 | 周淑芬; 邱慶豐; 王淑珍; 張竣凱; | 書刊名 | 嘉南學報. 科技類 |
卷 期 | 30 民93.12 |
頁 次 | 頁101-108 |
分類號 | 418.8 |
關鍵詞 | 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; |
語 文 | 中文(Chinese) |
中文摘要 | 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10[8fb6]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(enzyme-linked immunosorbent assay, indrect 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., ATTCC 13313),腸道出血型大腸桿菌(enterotoxigenic E. coli, EHEC, E. coli O157:H7, BCRC 13086)以及腸道毒素型大腸桿菌(enterohemorrhagic 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10[8fb6] CFU/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。 |
英文摘要 | 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. coil (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 10[8fb6] CFU/ml enterobacteria. |
本系統中英文摘要資訊取自各篇刊載內容。