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使用聚乙二醇6000从蛋黄中提取无污染物免疫球蛋白Y抗体的优化方法。

Optimized method for the extraction of contaminant-free IgY antibodies from egg yolk using PEG 6000.

作者信息

Madera-Contreras Andrea Michelle, Solano-Texta Roberto, Cisneros-Sarabia Alondra, Bautista-Santos Isabel, Vences-Velázquez Guillermina, Vences-Velázquez Amalia, Cortés-Sarabia Karen

机构信息

Facultad de Ciencias Químico-Biológicas of the Universidad Autónoma de Guerrero, Mexico.

出版信息

MethodsX. 2022 Oct 6;9:101874. doi: 10.1016/j.mex.2022.101874. eCollection 2022.

Abstract

Hens are oviparous vertebrates and produce IgY antibodies, which is the main type of immunoglobulin in the egg yolk, and high concentrations can be obtained by using a simple method that does not require sophisticated equipment and reagents. The Polyethylene Glycol 6000 method allows the removal of lipids and the precipitation of IgY in two days with an approximated purity of around 80%, however during the original protocol other contaminant proteins can be precipitated. To overcome the issue of contamination with other proteins and extraction time, we optimized the previously method described by Pauly et al. (2011) by adding some changes that improved the aforementioned problems. • Our protocol is customized by the addition of one more filtration step or one more step with PEG 6000 at 3.5% to avoid the contamination with lipids. • Additionally, the changes in the type of agitation, centrifugation and the skip of dialysis make the method more accessible for all the laboratories. • In summary, these modifications serve to enhance the purity, reduce the time for IgY extraction from egg yolk and make it more accessible for every basic research laboratory.

摘要

母鸡是卵生脊椎动物,能产生卵黄免疫球蛋白(IgY)抗体,这是蛋黄中的主要免疫球蛋白类型,通过一种不需要复杂设备和试剂的简单方法就能获得高浓度的IgY。聚乙二醇6000法可在两天内去除脂质并沉淀IgY,纯度约为80%,然而在原始方案中,其他污染蛋白也可能被沉淀。为了克服其他蛋白污染和提取时间的问题,我们对保利等人(2011年)描述的先前方法进行了优化,增加了一些改进上述问题的改动。• 我们的方案通过增加一个过滤步骤或一个3.5%聚乙二醇6000的步骤进行定制,以避免脂质污染。• 此外,搅拌类型、离心的改变以及透析步骤的省略使该方法对所有实验室来说更容易操作。• 总之,这些改进有助于提高纯度,减少从蛋黄中提取IgY的时间,并使每个基础研究实验室都更容易操作。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/861f/9563561/399bf454433f/ga1.jpg

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