三种鸡形目雉科禽类卵黄抗体的纯化及二抗的制备

Purification of IgYs from the Egg Yolk of Three Phasianidae Birds and Preparation of Rabbit Anti-IgY Antibodies

目的纯化三种鸡形目雉科的禽类孔雀、鹌鹑、贵妃鸡的卵黄抗体IgY,并且免疫家兔制备抗血清。方法采用了水稀释法,HiTrap IgYPurification HP免疫亲和层析法和硫酸铵沉淀法纯化IgY。免疫家兔制备抗血清,免疫双扩散法测定效价。Protein-A亲和纯化兔抗IgY血清IgG。结果经亲和纯化和盐析纯化,得到了孔雀、鹌鹑、贵妃鸡的卵黄抗体IgY,经SDS-PAGE检测为电泳纯,孔雀、鹌鹑、贵妃鸡的卵黄抗体IgY的相对分子质量约为180×103。免疫后经Protein-A亲和纯化后获得了兔抗IgY的IgG。结论证实了孔雀、鹌鹑、贵妃鸡的卵黄抗体IgY的存在及其特性。雉科鸡形目禽类卵黄抗体的纯化方法相似,可以推广到鸡形目其它禽类的卵黄抗体的纯化中,获得的抗血清可以进一步进行标记和今后抗原的检测。

Objective To purify the IgYs from the egg yolk of peacock and quail and ladyhood-chicken. Rabbits were immunized with the purified IgYs. Methods We employed the methods of water dilution affinity chromatograph and salt precipitation. The antiserum were characterized by double immunodiffusion （DID）. Results The purified products were identified by SDS-PAGE. The molecular weight of egg yolk antibodies of peacock and quail and ladyhood-chicken was 180 KD. The titer of the antiserum is high. Conclusion The purification methods for IgYs from peacock, quail and ladyhood-chicken were established.