特异性抗肠出血性大肠杆菌O157:H7内毒素鸡卵黄抗体的制备  

Preparation Study on Specific Egg Yolk Antibody against Endotoxin of Escherichia coli O157:H7

作　　者：宋宏新[1] 刘晓阳[1] 程军[1] 李书丰[2] 

机构地区：[1]陕西科技大学生命科学与工程学院,陕西咸阳712081 [2]德州职业技术学院,山东德州253034

出　　处：《食品科学》2007年第1期164-167,共4页Food Science

摘　　要：目的:比较研究LPS和O-SP的免疫原性,制备出特异性抗内毒素(LPS)鸡卵黄免疫球蛋白(eggyolkimmunoglobulin,IgY),用于对E.coliO157:H7的免疫预防及检测。方法:分别用灭活E.coliO157:H7菌体、不同浓度内毒素(LPS)及O-特异性多糖链(O-SP)与不完全弗氏佐剂充分乳化后作抗原免疫临产蛋母鸡,以水稀释法从卵黄中提取抗体,阴离子交换色谱法实现对抗体的纯化,间接ELISA及双向免疫扩散检测抗体产生效价与活性。结果与结论:所制内毒素(LPS)和O-SP均有较好的免疫原性,刺激鸡体后可产生高效价抗体,灭活菌体、600μg/mlLPS、1200μg/mlLPS、2000μg/mlLPS、O-SP抗体产生效价最高可分别达1:32000、1:28000、1:32000、1:12000、1:40000。结合离子交换色谱,用0.185mol/L的离子强度可实现一步洗脱纯化抗体,制备出高纯度、高活性、特异性强的鸡卵黄免疫球蛋白IgY,为大肠杆菌O157:H7的感染防治提供了可靠的保证,在此实验基础之上可进一步探讨应用特异性IgY对大肠杆菌O157:H7的检测。Objective： To produce specific chicken egg yolk immunoglobulin against ）, used for preventing and detecting Ecoli O157：H7 in comparison with the immunity of LPS and O-specific polysaccharide （O-SP）. Methods： Hens, 25-week old, were immunized with E. coli O157 ：H7 LPS O-SP respectively after being emulsified with incomplete Freuds＇ adjuvant. By a simple method of water dilute of protein IgY to separate the antibody from egg yolk, purified by ion-exchange chromatograph, the IgY detected by SDS-PAGE and indirect-ELISA, and its immunologic competence was determined by double immunodiffusion. Results and Conclusion：The IgY liters of E.coli O157：H7 are obtained with 60μg/ml LPS, 1200μg/ml LPS and 200μg/ml LPS, as O-SP respeclively 1：32000, 1：28000, 1：32000, 1：12000 and 1：40000. It is purified by ion exchanging with 0.185mol/L THS washing. The immunity of lipopolysaccharide and O-SP is strong. The IgY anti-OSP produced by immunizing hens shows high liter, high purification and well specificity by chicken egg yolk immunoglobulin. The results provide scientific basis for preventing and infection curing by O157：H7 and further ways exploring of detecting O157：H7 with specific anti-LPS IgY produced from hens.

关 键 词：E.COLI O157:H7 防治 内毒素 鸡卵黄抗体 制备 

分 类 号：Q939.91[生物学—微生物学] Q503