霍亂毒素多株抗體之製備與純化

CNU IR > Chna Nan Annual Bulletin > No.30 (2004) > Item 310902800/21334

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Title:	霍亂毒素多株抗體之製備與純化 Production and Purification of Polyclonal Antibodies Against Cholera Toxin
Authors:	周淑芬邱慶豐王淑珍張竣凱
Contributors:	生物科技系食品科技系
Keywords:	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
Date:	2004
Issue Date:	2009-06-01 11:41:37 (UTC+8)
Abstract:	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.
Relation:	嘉南學報(科技類)30期：p.101-108
Appears in Collections:	[Chna Nan Annual Bulletin] No.30 (2004) [Dept. of Biotechnology (including master's program)] Periodical Articles [Dept. of Food Science & Technology] Periodical Articles

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