抗白念珠菌鸡卵黄抗体的制备及温度对其效价的影响

目的:制备抗白念珠菌鸡卵黄抗体(IgY)并检测其生物学性能.方法:白念珠菌标准菌株10231灭活后作为抗原免疫25周龄产蛋海蓝鸡,从水溶法提取鸡卵黄IgY抗体.用考马斯亮蓝染色法测定IgY抗体提取液的蛋白浓度.用Tris-tricine电泳测定提取蛋白的相对分子质量、纯度,Western blot确定蛋白来源.以免疫组织化学检验抗白念珠菌IgY抗体与白念珠菌结合的特异性.ELISA测定抗白念珠菌IgY抗体的抗菌效价.结果:IgY抗体液中蛋白浓度为7.9 g/L,IgY抗体的纯度达到91.5%.提取的IgY抗体与鸡血清和正常鸡IgY抗体具有同源性.提取的IgY抗体可与白念珠菌发生特异性结合. ELISA测定抗白念珠菌IgY抗体的效价达到1:12 000.结论:研究得到的IgY,产量和纯度高,效价好,体温条件下48 h效价没有降低.在体外能够特异性地与白念珠菌结合.

Preparation of chicken egg yolk antibody against Candida albicans and effect of temperature on its titer

Objective: To prepare chicken egg yolk antibody(IgY) against Candida albicans and to study its biological properties in the interest of its further application.Methods: Ten 25 weeks old laying hens were immunized with Candida albicans.IgY was isolated from the yolks of eggs by the water dilution(WD) method.Its yield,purity,and molecular weight were measured by Bradford method and Tris-tricine gel electrophoresis,respectively.The antigenicity of IgY was determined with Western-blot method.The specific combination of IgY and Candida albicans was assayed with immunohistochemistry method.The activity of IgY was evaluated by enzyme-linked immunosorbent assay(ELISA).Results: The yield and purity of IgY were 7.9 g/L and 91.5%,respectively.It had the same molecular weight and antigenicity in comparison with IgY of chicken serum and normal chicken IgY.It could specifically combine with Candida albicans.Its titer was 1:12 000.Conclusion:The production and purity of IgY were high.Its titer was good and did not decrease under a body temperature condition after 48 h.The IgY antibody had a combination capacity of specificity for Candida albicans in vitro.