抗轮状病毒IgY和Fab’的分离与纯化

Preparation and Purification of IgY and Fab' Against Human Rotavirus

抗轮状病毒IgY可用两步盐析结合凝胶过滤从蛋黄中分离出来,用SDS-PAGE检测其纯度可达到95%以上。纯的IgY经胃蛋白酶分解得到的抗体片断(Fab),经SDS-PAGE和MALDI质谱法测定,其纯度达到99%以上。结果表明,所设计的分离抗轮状病毒IgY和Fab的方法简单、有效。经ELISA法检测,Fab的活性仍保持在IgY原始活性的70%以上。

Human Rotavirus （HRV） were used as antigen to immunize laying chickens and anti-HRV IgY was isolated from the yolk of eggs The purity was over 95 % by antibody fragment Fab＇ was methods, and its purity was isolating hen antibodies （IgY activity of IgY determined by using two salting-out steps followed by one-gel fihration chromatography methods. SDS-PAGE analysis. The purified IgY was digested with pepsin and the smaller obtained. The purified Fab＇ was characterized by SDS-PAGE and MALDI MS over 99%. The results indicate that the methods designed in this paper for and Fab＇） are simple and effective, the retained Fab＇ is over 70% of the initial the ELISA method.