Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13787
標題: 胸膜肺炎放線桿菌溶血素單株抗體之製作
Monoclonal Antibody Preparation against the Hemolysin of Actinobacillus Pleuropneumoniae
作者: 梁嘉君
Liang, Judging
關鍵字: Actinobacillus
胸膜肺炎放線桿菌
Hemolysin
Cytolysin
Haemophilus
Monoclonal antibody
Hybridoma.
溶血素
單株抗體
胸膜肺炎嗜血桿菌
溶細胞素
融合瘤。
出版社: 獸醫學系
摘要: 本實驗之目的在於製備胸膜肺炎放線桿菌第一血清型溶血素(HylI)的單株 抗體 (MAbs),並且以小白鼠為實驗動物注射MAbs後再接種胸膜肺炎放線 桿菌第一血清型細菌,觀察對小白鼠的保護效果。在單株抗體的製備方面 ,經過4 次重複免疫純化HylI 而有良好抗體反應的 BALB/c小白鼠,取脾 臟細胞和骨髓瘤細胞融合,結果在880孔中有24孔於酵素連結免疫吸附法 中呈強陽性反應,經 3次單株化後,得到9株分泌抗體能力穩定的融合瘤 ,其中有2株具有抗溶血作用,以其注射小白鼠腹腔24小時及鼻腔1小時後 接種胸膜肺炎放線桿菌第一血清型細菌,比用全菌免疫者,小白鼠的存活 率較高,為8:5。 對照組(免疫PBS)幾乎全數死亡;攻擊之活菌,可由斃 死之小白鼠肺臟回收。
The objective of this study is to prepare monoclonal antibodies (MAbs) against the hemolysin of Actinobacillus pleuropneumoniae (A.P.) serotype 1 (HylI). These MAbs were then used in the injection of mice, to assess their protection activity against A.P. challenge. The spleens of BALB/c mice had been immunized 4 times with purified HylI and showed very strong antibody response.These spleen cells were then fused with P3x myeloma cells. When screening test for specific antibodies production using the ELISA test, 24 out of 880 wells showed strong positive reactions. Subsequently 3 consecutive subclonings had established 9 strains of hybridoma with stable activities of MAbs secretion. Two strains among them with resist-hemolytic reactions injected mice. When comparing those immunized mice with those have been immunized by the bacteria, the survival ratio was 8:5 after the A.P. challenge. Whereas the contral groups (immunized with PBS) were killed after the challenge. Those challenge bacteria could be recovered from the lungs of dead mice.
URI: http://hdl.handle.net/11455/13787
Appears in Collections:獸醫學系所

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