抗轮状病毒IgY和Fab的研制

Preparation of IgY and Fab against HRV

目的对抗轮状病毒(RV)IgY和胃蛋白酶水解片断Fab进行分离与纯化。方法免疫鸡得到抗HRVIgY,用两步盐析结合凝胶过滤将其从蛋黄中分离出来,纯的IgY经胃蛋白酶水解得抗体片断Fab。结果抗HRVIgY用SDS-PAGE检测纯度可达到95%以上。抗轮状病毒(RV)IgY和抗体片断Fab经SDS-PAGE和MALDIMS法测定,其纯度达到99%以上,经ELISA法检测,Fab的活性保持在Igy原始活性的70%以上。结论我们所设计的分离和纯化抗HRVIgY和Fab的方法简单、有效。

Objective To isolate and purify the IgY and Fab against human rotavirus (HRV). Methods HRV were used as antigen to immunize laying chickens. anti-HRV IgY was isolated from the egg yolk of these chickens using two-step salting-out and one-gel filtration chromatography. The purified IgY was digested with pepsin and the smaller antibody fragment Fab was obtained. Results The purity of anti-HRV IgY was over 95% detected by SDS-PAGE, while the purities of anti-HRV IgY and the purified Fab were both over 99% by detected by SDS-PAGE and MALDI MS methods. The antigen-binding activities of IgY and Fab were comparatively analyzed by the ELISA, which showed Fab retained over 70% of the initial activity of IgY. Conclusion The designed methods for isolating hen antibodies (IgY and Fab) are simple and effective.