What is it about?
Two-dimensional gel electrophoresis (2-DE) is a technique that has been widely applied in a variety of proteomics studies. It is capable of resolving complex protein mixtures into individual protein spots based on their isoelectric point and molecular weight, enabling large-scale analysis of protein expression patterns for deciphering their changes in different biological conditions. 2-DE is a powerful tool that empowers researchers to perform differential qualitative and quantitative proteome analysis and is particularly advantageous for characterizing protein isoforms and post-translationally modified proteins. Despite its popularity as the workhorse for proteomics in the past few decades, it has been gradually displaced by the more sophisticated and high-performance mass spectrometry-based methods. However, there are several variations of the 2-DE technique that have emerged as promising approaches that shine new light on specific niches that 2-DE could still contribute. In this review, we first provide an overview of the applications of 2-DE, its merits and pitfalls in the current proteomic research arena, followed by a discussion on several alternative approaches for potential future applications.
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Why is it important?
• Review of current trends of 2-DE applications in the field of proteomics. • Strengths and shortcomings of 2-DE technique. • Innovative adaptations of 2-DE for alternative applications and future prospects.
Perspectives
Since the birth of proteomics, 2-DE is the workhorse to characterize protein profiles of various biological samples. It represents an affordable and useful technique for high-resolution separation of complex protein mixtures that enables quantitative differential protein expression profiling. However, the 2-DE technique also faces several technical challenges such as sensitivity and reproducibility, as well as being laborious that limit its practical use. Consequently, further technological improvements and advances of methodologies are desirable to overcome these limitations. Even though the field of proteomic today has been largely dominated by the state-of-the-art mass spectrometry-based technique, it is clear that 2-DE still plays an important role in proteomic research. Among the unique strengths of 2-DE are that it delivers valuable information of the proteome, including the molecular weight, isoelectric point, abundance level, the presence of protein modifications, and isoforms that makes it irreplaceable by any other current approaches. Furthermore, it is interesting to note that the incorporation of new ideas and modifications to the existing 2-DE technique has led to the development of several promising alternative techniques that could expand its applications for proteomic analysis. Hence, we envision that 2-DE will continue to serve as a powerful and complementary tool for interrogating the proteome profiles and further contribute to our biological understanding.
Dr. Neda Saraygord-Afshari
Iran University of Medical Sciences
Read the Original
This page is a summary of: The evolution of two-dimensional gel electrophoresis - from proteomics to emerging alternative applications, Journal of Chromatography A, March 2020, Elsevier,
DOI: 10.1016/j.chroma.2019.460763.
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