ReviewTwo-dimensional gel electrophoresis in proteomics: Past, present and future
Graphical abstract
This review article starts with the birth of 2D electrophoresis, investigates how it has been instrumental to the birth of proteomics, and examines its position in the current proteomics landscape.
Section snippets
From mute star maps to protein buzz: two-dimensional electrophoresis and the birth of proteomics
SDS electrophoresis in its modern form was introduced in the early 70s [1] and soon became very widely used by protein biochemists. However, analysis of complex cellular extracts by this method made obvious the fact that the resolution was far from being sufficient to separate the many protein components of such extracts. To increase the resolution to a sufficient extent, it was necessary to couple two independent separations. At that time (and this is still the case) it was obvious that the
The take-off of proteomics
This possibility to be able to analyze spot of interest coming from 2D gels was a crucial change of paradigm, and the real start of proteomics. At those times where no complete genome was published yet, Edman sequencing provided enough information to look for homologs, or to devise oligonucleotides for screening DNA libraries. It was also the starting of brief golden age of 2D gel images databases [43], [44], [45], [46], [47]. To be useful to the community, 2D gel image databases must contain
Touching the limits of 2D electrophoresis, and the creation of alternate methods
Because of this intensive and worldwide effort using 2D electrophoresis and mass spectrometry as the core tool, many data were accumulated and analyzed, and it soon became obvious that it was always the same types of proteins that were found again and again, and the same types that were always missing, i.e. the low abundance and the hydrophobic proteins [61], [62].
In order to improve the resolution of hydrophobic proteins, many efforts were devoted to improve protein solubilization under the
Using the strengths of 2D gels in modern proteomic research
In this section, we will try to sort out the major strengths of 2D gels and to exemplify how they can be used for the benefit of proteomic research.
Concluding remarks. What future for 2D gels in proteomics
From all of the above, it is tempting to try to predict the future uses of 2D electrophoresis in the future, although this prediction exercise is always difficult and risky. It can be analyzed, however, that in the present proteomics landscape, 2D electrophoresis has two main drawbacks and three main advantages. The two main drawbacks are its very low efficiency (to say the least), in the analysis of hydrophobic proteins, and its high sensitivity to the dynamic range and quantitative
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