Suppr超能文献

一种新颖、经济高效的鸡卵免疫球蛋白 Y 纯化方法。

A novel, cost-effective and efficient chicken egg IgY purification procedure.

机构信息

Department of Chemistry & Biomolecular Sciences, Macquarie University, NSW, 2109, Australia.

出版信息

J Immunol Methods. 2012 Jun 29;380(1-2):73-6. doi: 10.1016/j.jim.2012.03.003. Epub 2012 Mar 29.

Abstract

Chicken IgY antibodies have been touted to be a superior alternative to mammalian antibodies for use in various immunological, molecular biology and proteomics applications for several reasons. These include, but are not limited to, improved specificity due to maximum phylogenetic distance between host and recipient, cost effectiveness in maintaining commercial numbers of hens, IgY yield and the use of non-invasive methods used to isolate IgY from eggs as opposed to blood. Despite this, the routine use of IgY-based methodologies in the laboratory is not widespread. One reason for this reluctance may be derived from the difficulties and expense of isolating IgY antibodies from egg yolk in sufficient yield, with high purity at a realistic reasonable price. Here, we describe an extremely cost-effective ($5USD per egg), rapid (within 5 h), efficient and optimised technique to isolate high yields (60 mg) of high purity (~80%) chicken IgY from egg yolks using the common plant gums pectin and κ-carrageenan in the presence of calcium chloride to delipidate egg yolk mixtures whilst maintaining IgY in solution and then ammonium sulphate to subsequently precipitate the resulting IgY antibodies to higher purity. Our data demonstrates that this technique results in a high yield and purity of IgY that is comparable (if not superior to) existing commercial IgY isolation kits. The method also allows the isolation of immunologically active IgY which can be used for further downstream immunotechnological processes. Furthermore, it can also be easily implemented in a standard well equipped laboratory, and may be scaled up to commercial quantities (i.e., thousands of eggs).

摘要

鸡 IgY 抗体因其在各种免疫学、分子生物学和蛋白质组学应用中的诸多优势而被吹捧为优于哺乳动物抗体的替代品。这些优势包括但不限于宿主和受体之间最大的进化距离导致的更高特异性、维持商业数量的母鸡的成本效益、IgY 的产量以及与从血液中分离 IgY 相比使用非侵入性方法从鸡蛋中分离 IgY。尽管如此,IgY 为基础的方法在实验室中的常规使用并不广泛。这种不情愿的原因之一可能源于从蛋黄中分离 IgY 抗体的困难和成本,需要以合理的价格获得足够的产量、高纯度。在这里,我们描述了一种极其经济高效(每个鸡蛋 5 美元)、快速(5 小时内)、高效且优化的技术,可使用常见的植物胶果胶和 κ-卡拉胶在氯化钙存在下从蛋黄中分离出高纯度(~80%)的高产量(60mg)鸡 IgY,同时使蛋黄混合物脱脂蛋白并保持 IgY 溶解,然后使用硫酸铵将所得 IgY 抗体沉淀至更高纯度。我们的数据表明,该技术可获得高产量和高纯度的 IgY,与现有的商业 IgY 分离试剂盒相当(如果不是优于)。该方法还允许分离具有免疫活性的 IgY,可用于进一步的下游免疫技术过程。此外,它还可以在标准设备齐全的实验室中轻松实施,并可扩展到商业规模(即数千个鸡蛋)。

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验