An optimized purification process for porcine gastric mucin with preservation of its native functional properties
An optimized purification process for porcine gastric mucin with preservation of its native functional properties
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DOI：
10.1039/c6ra07424c
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发表时间：
2016-01-01
期刊：
影响因子：
3.9
通讯作者：
Berensmeier, Sonja
中科院分区：
文献类型：
--
作者：
Schoemig, Veronika J.;Kaesdorf, Benjamin T.;Berensmeier, Sonja
纯化的胃粘蛋白目前用于广泛的应用，例如作为天然粘液的模型系统，作为润滑剂或抗病毒/抗菌补充剂。然而，市售的猪胃粘蛋白（PGM）不显示凝胶形成特性，并且仅显示大大降低的抗病毒/抗细菌活性。因此，我们建立了一个强大的纯化工艺的PGM，保持其所需的性质，如润滑性，凝胶形成和分子的选择性结合。我们优化了工艺的收率和生产力，并评估了不同的缓冲液条件对粘蛋白质量的影响。引入并优化使用IOOkDa膜的错流过滤以在尺寸排阻色谱之前预浓缩粘蛋白溶液。渗滤后电导率小于100 μ S cm（1）被认为是凝胶形成的关键。优化工艺的粘蛋白得率为66%。放大导致每mL粗粘液每小时0.15mg纯化粘蛋白的生产率。总的来说，大约。每只猪胃可纯化65 mg粘蛋白。摩擦学研究，流变学测量和共定位实验证实了保留功能的纯化粘蛋白的润滑性，凝胶形成和带电分子的结合相互作用，分别。
Purified gastric mucins are currently used for a wide range of applications e.g. as a model system for native mucus, as lubricants or antiviral/antibacterial supplements. However, commercially available porcine gastric mucins (PGM) do not exhibit gel-forming properties and show only greatly reduced anti-viral/anti-bacterial activity. Thus, we established a robust purification process for PGM, maintaining its desired properties such as lubricity, gel formation and the selective binding of molecules. We optimized the process in terms of yield and productivity and evaluated the influence of different buffer conditions on mucin quality. Cross-flow filtration using 100 kDa membranes was introduced and optimized to pre-concentrate the mucin solution prior to size exclusion chromatography. A conductivity of less than 100 mu S cm (1) after diafiltration was found to be crucial for gel formation. The mucin yield of the optimized process was 66%. The scale-up resulted in a productivity of 0.15 mg purified mucin per mL crude mucus an hour. In total, approx. 65 mg mucin could be purified from one pig stomach. Tribological studies, rheological measurements and co-localization experiments confirmed the retained functionality of purified mucin in terms of lubricity, gel formation and binding interactions with charged molecules, respectively.