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[鸡卵黄免疫球蛋白的制备及GoldMag载体偶联制备的鸡免疫球蛋白对血浆部分高丰度蛋白的去除]

[Preparation of hen egg yolk immunoglobulin and partial plasma high abundant protein depletion by prepared chicken immunoglobulin coupled on GoldMag carrier].

作者信息

Lin Fang, Huang Jian-Xin, Chen Chao, Liu Jia, Cui Ya-Li

机构信息

Life Science College, Northwest University, Xi'an 710069, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jul;24(7):706-9.

Abstract

AIM

To prepare specific egg yolk immunoglobulin (IgY) against HSA and IgG and to make HSA and IgG depletion research in human plasma by binding it to the surface of GoldMag.

METHODS

Laying Roman hens immunized with different antigens, such as HSA, IgG and the mixtures of these proteins were used in the experiments. The best condition of isolating the IgY antibody was studied. The titer and purity of the antibody were examined by indirect ELISA and SDS-PAGE respectively. Then the specific IgY was bound to the surface of GoldMag for HSA and IgG depletion research.

RESULTS

60-120 days after immunization, the titer of the antibody in the plasma against different antigens reached 1:15 000-1:25 000; while the titer in the egg reached 1:10 000-1:25 000. The purity was over 98%. Most of HSA and IgG were depleted from human plasma by IgY connected to GoldMag.

CONCLUSION

HSA and IgG can be used as antigens to immunize laying hens to produce IgY with high titer and purity. IgY which is bound to the GoldMag carrier can be used to deplete of the relevant HSA and IgG from human plasma effectively. As a new method of studying proteome of plasma, it has some practical applications.

摘要

目的

制备针对人血清白蛋白(HSA)和免疫球蛋白G(IgG)的特异性蛋黄免疫球蛋白(IgY),并通过将其结合到磁珠表面实现人血浆中HSA和IgG的去除研究。

方法

使用经不同抗原(如HSA、IgG及其蛋白混合物)免疫的罗曼蛋鸡进行实验。研究分离IgY抗体的最佳条件。分别通过间接酶联免疫吸附测定(ELISA)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测抗体的效价和纯度。然后将特异性IgY结合到磁珠表面进行HSA和IgG去除研究。

结果

免疫后60 - 120天,血浆中针对不同抗原的抗体效价达到1:15 000 - 1:25 000;而蛋黄中的效价达到1:10 000 - 1:25 000。纯度超过98%。与磁珠相连的IgY能有效去除人血浆中的大部分HSA和IgG。

结论

HSA和IgG可作为抗原免疫蛋鸡以产生高效价和高纯度的IgY。结合到磁珠载体上的IgY可有效去除人血浆中相关的HSA和IgG。作为一种研究血浆蛋白质组的新方法,具有一定的实际应用价值。

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