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电泳在蛋白质定量中的适用性

On the Applicability of Electrophoresis for Protein Quantification.

作者信息

Dome Karina, Akimenko Zoya, Bychkov Aleksey, Kalambet Yuri, Lomovsky Oleg

机构信息

Institute of Solid State Chemistry and Mechanochemistry, Siberian Branch, Russian Academy of Sciences, 630128 Novosibirsk, Russia.

Department of Business, Novosibirsk State Technical University, 630073 Novosibirsk, Russia.

出版信息

Polymers (Basel). 2021 Nov 17;13(22):3971. doi: 10.3390/polym13223971.

Abstract

Polyacrylamide gel electrophoresis is widely used for studying proteins and protein-containing objects. However, it is employed most frequently as a qualitative method rather than a quantitative one. This paper shows the feasibility of routine digital image acquisition and mathematical processing of electropherograms for protein quantification when using vertical gel electrophoresis and Chrom & Spec software. Both the well-studied model protein molecules (bovine serum albumin) and more complex real-world protein-based products (casein-containing isolate for sports nutrition), which were subjected to mechanical activation in a planetary ball mill to obtain samples characterized by different protein denaturation degrees, were used as study objects. Protein quantification in the mechanically activated samples was carried out. The degree of destruction of individual protein was shown to be higher compared to that of the protein-containing mixture after mechanical treatment for an identical amount of time. The methodological approach used in this study can serve as guidance for other researchers who would like to use electrophoresis for protein quantification both in individual form and in protein mixtures. The findings prove that photographic imaging of gels followed by mathematical data processing can be applied for analyzing the electrophoretic data as an affordable, convenient and quick tool.

摘要

聚丙烯酰胺凝胶电泳广泛应用于蛋白质及含蛋白质物质的研究。然而,它最常被用作定性方法而非定量方法。本文展示了在使用垂直凝胶电泳和Chrom & Spec软件时,对电泳图谱进行常规数字图像采集和数学处理以实现蛋白质定量的可行性。研究对象既包括经过充分研究的模型蛋白质分子(牛血清白蛋白),也包括更为复杂的基于现实世界蛋白质的产品(用于运动营养的含酪蛋白分离物),这些产品在行星式球磨机中进行机械活化以获得具有不同蛋白质变性程度特征的样品。对机械活化样品进行了蛋白质定量。结果表明,在相同的机械处理时间后,单个蛋白质的破坏程度高于含蛋白质混合物。本研究中使用的方法学方法可为其他希望将电泳用于单个蛋白质和蛋白质混合物定量的研究人员提供指导。研究结果证明,凝胶的摄影成像及随后的数学数据处理可作为一种经济实惠、方便快捷的工具用于分析电泳数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17de/8625784/6dbb75c3e85a/polymers-13-03971-g001.jpg

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