摘要
目的探索有效的纯化大雁卵黄抗体及高效价二抗的方法。方法应用了PEG沉淀,阴离子交换柱,硫酸铵分级沉淀纯化,免疫制备二抗,免疫电泳和免疫双扩散法及western blotting进行效价的测定。结果改良后的硫酸铵沉淀法的纯度远远高于PEG沉淀,相对于离子交换法而言更加简单方便,经济省时。从免疫电泳看已达到了电泳纯,大雁卵黄IgY的相对分子质量为180×103,重链约为66×103,轻链约为24×103。免疫获得的二抗有较强的免疫活性。结论用改良的硫酸铵沉淀法纯化大雁卵黄抗体,效率和纯度显著提高,值得推广。本实验为大雁卵黄抗体及其二抗的应用奠定了基础,也为雁形目其他种类卵黄抗体的纯化提供了参考。
Objective To isolate lgY from the wild goose egg yolk and produce antiserum. Methods We employed the method of PEG precipitation, ion exchange chromatography, and sequential ammonium sulfate precipitation to purify the wild goose lgY. Rabbits were inoculated with the purified lgY and the antiserum was identified by double immunodiffusion and western blotting. Results The improved ammonium sulfate precipitation method is better and convenient than PEG precipitation and ion exchange chromatography. SDS-PAGE indicated that the molecular weight of wild goose IgY is 180 × 10^3 consisted with heavy chain of 66 × 10^3 and light chain of 24 × 10^3 . The polyclonal rabbit anti-wild goose lgY antiserum was produced with a titer of 1 ： 32. Conclusion Ammonium sulfate precipitation method was established for wild goose lgY.
出处
《中国比较医学杂志》
CAS
2007年第11期625-628,共4页
Chinese Journal of Comparative Medicine
基金
社会公益研究专项(2005DIA1J095)
关键词
大雁
IGY
硫酸铵沉淀
Wild goose
lgY
Ammonium sulfate precipitation
