摘要
目的 制备特异抗内毒素鸡蛋黄免疫球蛋白 (Eggyolkimmunoglobulin ,IgY) ,探索防治内毒素血症的新途径。方法 用大肠杆菌J5株、内毒素 (Lipopolysaccharide,LPS)及类脂A(LipidA)作抗原免疫 2 5周龄Roman鸡 ,改良水溶法提取抗体 ,进行紫外分光光度计检测、SDS PAGE及Westernblot免疫印迹分析 ,通过酶联染色反应检测其免疫学活性。结果 大肠杆菌J5株、LPS及LipidA抗体含量分别为 11.4、9.2、9.3mg mL蛋黄液 ,质量分数分别为 92 .3%、87.13%、90 .4 % ,分子质量为180 0 0 0u ,初步鉴定其对内毒素具有特异性结合作用。结论 用大肠杆菌J5株、LPS及LipidA免疫鸡制备的IgY效价高、特异性强、产量大 。
ObjectiveTo produce specific chicken egg yolk immunogl ob ulin against lipopolysaccharide(LPS) in order to explore a new way of prevention and cure of endotoximia.MethodsRoman hens , 25 week old ,wer e immunized with Escherichia coli J5 , lipopolysaccharide and lipid A respec tively. By a simple method of water dilution protein purification IgY was separa ted f rom egg yolk . IgY was detected by spectrophotometer and analyzed by SDS PAGE a nd western blot, and its immunologic competence was determined by enzyme labelin g stai ning reaction.Results IgY from Escherichia coli J5 , lipo polysaccharide and lipid A was respectively found 11.4 , 9.2 , 9.3 mg/mL egg yo lk and 92.3% , 87.13% , 90.4% purity and its molecular weight was 180 000 u. conclusionIgY from Escheric hia coli J5 , lipo polysaccharide and lipid A against LPS produced by immunizing Roman hens had hig h titer ,well specificity and large production. It would be a new method of prev ention and cure of endotoximia.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2003年第1期66-68,共3页
Immunological Journal
基金
中关村医药科技创新基金资助项目 (0 2 0 0 10 )
