论文部分内容阅读
目的制备高效价特异性的抗沙门氏菌IgY抗体,并对其基本特性进行分析和鉴定。方法以沙门氏菌作为抗原免疫产蛋的京白鸡,待抗体产生后,经过50%饱和硫酸铵盐析、透析、以及聚乙二醇浓缩等步骤纯化抗体。蛋白质定量后,再通过SDS-PAGE和ELISA等方法对该抗体进行分析鉴定。结果经抗原免疫的母鸡,10d左右就可从其所产蛋的卵黄中检测出特异性IgY抗体,抗体量在30d左右达到高峰,经纯化的抗体通过还原性SDS-PAGE显示两条带,其分子量约为67kD和28kD,抗体纯度可达92.1%。结论成功制备了抗沙门氏菌IgY抗体,实验结果表明IgY抗体是一种产量高,制备简便的新型抗体,并且高效价抗体持续时间较长,其特异性也较好。
Objective To prepare high titer anti-Salmonella IgY antibody and to analyze and identify its basic properties. Methods The Jing-white chicken was immunized with Salmonella as an antigen. After antibody production, the antibody was purified by 50% saturated ammonium sulfate salting-out, dialysis, and polyethylene glycol concentration. After the protein was quantified, the antibody was analyzed and identified by SDS-PAGE and ELISA. Results The specific immunoglobulin A (IgY) antibody was detected in the egg yolk of the hens immunized by the antigen about 10 days after the immunization. The antibody level peaked at about 30 days. The purified antibody showed two bands by reducing SDS-PAGE. The molecular weight of about 67kD and 28kD, antibody purity up to 92.1%. Conclusion The anti-Salmonella IgY antibody was successfully prepared. The results showed that the IgY antibody is a new antibody with high yield and easy preparation, and the high titer antibody has a longer duration and better specificity.