Comparison of five procedures for the purification of insoluble elastin
Comparison of five procedures for the purification of insoluble elastin
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DOI：
10.1016/s0142-9612(00)00383-5
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发表时间：
2001-07-01
期刊：
影响因子：
14
通讯作者：
van Kuppevelt, TH
中科院分区：
文献类型：
--
作者：
Daamen, WF;Hafmans, T;van Kuppevelt, TH
弹性蛋白是不溶性的。高度交联的蛋白质。为肺、主动脉和韧带等器官提供弹性。尽管弹性蛋白具有显著的机械性能，但它作为生物材料的用途很少。从弹性纤维中纯化完整的弹性蛋白是一个主要的挑战，其中包括弹性蛋白和微纤维的紧密交织。不溶性弹性蛋白制剂倾向于钙化，这可能是由于钙结合微纤维（例如，胶原蛋白）。在这项研究中，弹性蛋白纯化马项韧带使用五种不同的程序。一种方法是基于用0.1 M NaOH处理，另一种是基于高压灭菌和用溴化氰处理。其他三种方法是基于提取步骤和酶消化的组合。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳评估制剂的纯度。氨基酸分析、亮保持免疫荧光和透射电子显微镜。涉及提取物/酶的程序与2-巯基乙醇和溴化氰的早期应用相结合，得到了高纯度的弹性蛋白制剂。电子显微镜分析表明，该制剂是：缺乏微纤维成分。因此，该过程是用于制备作为组织工程生物材料的不溶性弹性蛋白的选择方法。(C)2001 Elsevier Science Ltd.保留所有权利。
Elastin is an insoluble. highly cross-linked protein. providing elasticity to organs like lung, aorta, and ligaments. Despite its remarkable mechanical properties, elastin has Found little use as a biomaterial. Purification of intact elastin from elastic fibres presents a major challenge, among others for the intimate interwoveness of elastin and microfibrils. Insoluble elastin preparations tend to calcify, which may be due to calcium-binding microfibrillar (e.g. fibrillin). In this study, elastin was purified from horse ligamentum nuchae using five different procedures. One procedure is based on treatment with 0.1 M NaOH, another on autoclaving and treatment with cyanogen bromide. Three other procedures are based on combinations of extraction steps and enzyme digestions. Purity of preparations was assessed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. amino acid analysis, bright held immunofluorescence and transmission electron microscopy. The procedure involving extracrions/enzymes combined with an early application of 2-mercaptoethanol and cyanogen bromide gives a highly pure elastin preparation. Electron microscopic analysis showed that this preparation is: devoid of microfibrillar components. This procedure is therefore the method of choice for preparation of insoluble elastin as a biomaterial for tissue engineering. (C) 2001 Elsevier Science Ltd. AH rights reserved.