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首页> 外文期刊>Protein Expression and Purification >Purification and characterization of osteopontin from human milk
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Purification and characterization of osteopontin from human milk

机译：人乳中骨桥蛋白的纯化与表征

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Osteopontin (OPN) is expressed in many organs and tissues and has different biological properties related to different molecular forms in respect to size and posttranslational modifications. However, a purification procedure for authentic intact OPN as well as fragments of OPN from an accessible biological source is missing. A four-step procedure was used to purify OPN from human milk, based on its crystal growth inhibitory activity, including anion exchange chromatography, the elimination of casein, hydroxyapatite chromatography, and negative affinity chromatography. Purified OPN was further separated into its different molecular forms by means of a two-step procedure, involving size exclusion chromatography and reverse phase chromatography. A rabbit polyclonal antibody was raised to purified intact OPN and high M-r OPN components; the immunoreactivity of both forms was almost equal when investigated by enzyme immunoassay (EIA). The procedures facilitate the purification of intact OPN and OPN fragments for purposes of standardization, preparation of monospecific antibodies, and functional studies. (C) 2003 Elsevier Science (USA). All rights reserved. [References: 30]
机译：骨桥蛋白（OPN）在许多器官和组织中表达，就大小和翻译后修饰而言，具有与不同分子形式相关的不同生物学特性。但是，缺少用于真实完整OPN以及来自可及生物来源的OPN片段的纯化程序。基于其晶体生长抑制活性，采用四步法从人乳中纯化OPN，包括阴离子交换色谱，酪蛋白消除，羟基磷灰石色谱和负亲和色谱。纯化的OPN通过两步法进一步分离成其不同的分子形式，包括尺寸排阻色谱法和反相色谱法。产生兔多克隆抗体以纯化纯化的完整OPN和高M-r OPN组分;当通过酶免疫分析法（EIA）研究时，两种形式的免疫反应性几乎相等。该程序有助于纯化完整的OPN和OPN片段，以实现标准化，制备单特异性抗体和进行功能研究的目的。 （C）2003 Elsevier Science（美国）。版权所有。 [参考：30]

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