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膜表面蛋白质沉积层的半定量、空间分辨分析。

Semi-quantitative, spatially resolved analysis of protein deposit layers on membrane surfaces.

作者信息

Hartinger Martin, Napiwotzki Jonas, Schmid Eva-Maria, Kurz Franziska, Kulozik Ulrich

机构信息

Chair of Food and Bioprocess Engineering, Technical University of Munich, Germany.

出版信息

MethodsX. 2019 Dec 19;7:100780. doi: 10.1016/j.mex.2019.100780. eCollection 2020.

DOI:10.1016/j.mex.2019.100780
PMID:31993341
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6974784/
Abstract

Fouling distinctly reduces the filtration performance of membranes. A characterization of the fouling in membranes, however, is difficult due to its spatial distribution. Currently applied methods for deposit layer analysis are rather complex or do not offer a spatial resolution. Knowledge of the spatial distribution, however, could be used to improve the design of membranes, modules, and spacers. Staining with Coomassie Brilliant Blue, related to the staining of PAGE gels, is a simple method to visualize and analyze the deposited proteins semi-quantitatively. We improved an existing staining technique for protein deposits on membranes by adding a calibration for the semi-quantitative analysis and optimizing the sample handling. The method provides a spatially resolved analysis of deposited proteins up to a concentration of 10 g m. Apart from staining, data processing is described in order to generate false colors or topographic images of deposits. Thus, the paper describes a simple method to assess and visualize the influence of module characteristics such as spacer design on the spatially resolved protein fouling of polymeric and ceramic membranes. Therefore, the method can contribute to the improvement of the module design and processing conditions with regard to the filtration performance. •Visualization of proteinaceous deposits on membranes•Spatially resolved quantification of proteinaceous deposits.

摘要

污垢明显会降低膜的过滤性能。然而，由于污垢的空间分布，对膜中污垢的表征很困难。目前应用的沉积层分析方法相当复杂，或者没有提供空间分辨率。然而，污垢空间分布的相关知识可用于改进膜、膜组件和间隔物的设计。与聚丙烯酰胺凝胶电泳（PAGE）凝胶染色相关的考马斯亮蓝染色，是一种简单的方法，可用于半定量地可视化和分析沉积的蛋白质。我们通过添加半定量分析的校准并优化样品处理，改进了现有的膜上蛋白质沉积物染色技术。该方法可对浓度高达10 g/m的沉积蛋白质进行空间分辨分析。除了染色，还描述了数据处理过程，以生成沉积物的伪彩色或地形图。因此，本文描述了一种简单的方法，用于评估和可视化诸如间隔物设计等膜组件特性对聚合物膜和陶瓷膜空间分辨蛋白质污垢的影响。因此，该方法有助于在过滤性能方面改进膜组件设计和处理条件。•膜上蛋白质沉积物的可视化•蛋白质沉积物的空间分辨定量分析

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/c3118095f03c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/6a9e58b5a57a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/896d9f595c63/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/7f0f91c5dcfb/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/d757619f976f/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/07742e5eea08/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/623c03b65de5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/81706bfef4a9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/c3118095f03c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/6a9e58b5a57a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/896d9f595c63/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/7f0f91c5dcfb/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/d757619f976f/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/07742e5eea08/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/623c03b65de5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/81706bfef4a9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d4f/6974784/c3118095f03c/gr7.jpg

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引用本文的文献

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本文引用的文献

1
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2
Development of an ultrasonic technique for in situ investigating the properties of deposited protein during crossflow ultrafiltration.一种用于原位研究错流超滤过程中沉积蛋白质特性的超声技术的开发。
J Colloid Interface Sci. 2005 Apr 1;284(1):228-38. doi: 10.1016/j.jcis.2004.09.072.
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Nomenclature of the proteins of cows' milk--sixth revision.
牛乳蛋白质命名法——第六版修订本
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Mechanism of dye response and interference in the Bradford protein assay.考马斯亮蓝蛋白测定法中染料反应及干扰的机制
Anal Biochem. 1985 Dec;151(2):369-74. doi: 10.1016/0003-2697(85)90190-3.
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Why does Coomassie Brilliant Blue R interact differently with different proteins? A partial answer.为什么考马斯亮蓝R与不同蛋白质的相互作用不同？一个部分答案。
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Quantitation of proteins bound to polyvinylidene difluoride membranes by elution of coomassie brilliant blue R-250.通过考马斯亮蓝R-250洗脱定量与聚偏二氟乙烯膜结合的蛋白质。
Anal Biochem. 1990 Sep;189(2):169-72. doi: 10.1016/0003-2697(90)90101-e.
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A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.一种利用蛋白质 - 染料结合原理对微克级蛋白质进行定量的快速灵敏方法。
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