Discarded free PEG-based assay for obtaining the modification extent of pegylated proteins
Discarded free PEG-based assay for obtaining the modification extent of pegylated proteins
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DOI：
10.1016/j.talanta.2006.04.010
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发表时间：
2007-01-15
期刊：
影响因子：
6.1
通讯作者：
Xiong, Yu Chun
中科院分区：
文献类型：
--
作者：
Gong, Xing Wen;Wei, Dong Zhi;Xiong, Yu Chun
游离聚乙二醇（PEG）是聚乙二醇化过程中产生的副产物，在纯化聚乙二醇化蛋白质时应将其除去。本文将其应用于聚乙二醇化蛋白质修饰程度的测定。该方法包括两个操作步骤。首先，通过CM-Sepharose FF从聚乙二醇化蛋白的粗反应混合物中分离游离的PEG。然后根据与氯化钡和碘溶液形成络合物来测定PEG。PEG的有效检测范围为0 ~ 7.5 μ g/ml。根据公式计算改性程度。该方法简便、灵敏，适用于所有的PEG衍生物。它最大的特点是不消耗蛋白质。(c)2006 Elsevier B. V.保留所有权利。
Free polyethylene glycol (PEG) is a byproduct produced during the process of pegylation and should be removed for the purification of pegylated proteins. In this paper, it was used to develop a new method for obtaining the modification extent of pegylated proteins. This method included two steps of operation. Firstly, the free PEG was separated from crude reaction mixture of pegylated proteins by CM-Sepharose FF. Then PEG was determined based on the formation of a complex with barium chloride and iodine solution. The effective detective range of PEG was 0-7.5 mu g/ml. The modification extent was calculated according to a formula. This method is simple, sensitive, and applicable to all of the PEG derivatives. The most distinctive aspect is that it does not consume proteins. (c) 2006 Elsevier B.V. All rights reserved.