摘要
目的研究抗内毒素卵黄免疫球蛋白(IgY)的活性片断Fab’,探讨防治内毒素血症的新途径。方法用内毒素(LPS)作为抗原免疫25周龄德国罗曼鸡,改良水溶法提取抗内毒素IgY,胃蛋白酶切后提取Fab’片断,光密度法测抗内毒素Fab’的浓度和含量、ELISA检测抗内毒素Fab’效价、SDS-聚丙烯酰胺凝胶电泳检测其分子量及纯度。结果抗内毒素Fab’含量为4.2mg/mL蛋黄液,效价为1∶51200,纯度为92%,相对分子质量为44000。结论抗内毒素Fab’产量大、效价高、特异性强。
Objective To investigate the Fab＇ in the chicken egg yolk antibody against lipopolysaccharide （LPS） for exploring a new way of preventing and curing endotoximia. Methods The purified LPS together with Freund＇s adjuvant was used as antigen to immunize Luoman hens of 25 weeks old and the eggs laid by these hens were collected. Yolk immunoglobulin （IgY） against LPS was isolated from yolk sac of eggs, and then purified by water extraction, salt precipitation with ammonium sulfate, and dialysis. Then, the Fab＇ was prepared by the digestion of the pepsin. Titers and immunological activities of the antibody fragment were measured by ELISA; the purity of the antibody fragment was determined by SDS-PAGE. Results The protein concentration of Fab＇ was 4.2 mg/mL egg yolk. The purity and relative molecular weight of the Fab＇ were about 92% and 44 000, respectively. Tne titer of the Fab＇ was 1：51 200 demonstrated by ELISA; the specificity of the Fab＇ was confirmed by the Western blotting. Conclusion The anti-LPS Fab＇ with high fiter, well specificity, and large production has prepared successfully, thus providing an impossible new way for [Key words]
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第2期219-221,共3页
Immunological Journal
基金
国家重点基础研究发展规划资助项目(2005CB522601)
