Properties &
Functions
- Two-dimensional
electrophoresis (2-D electrophoresis) is a powerful and widely used
method for the analysis of complex protein mixtures extracted from
cells, tissues, or other biological samples.
- Two-dimensional
electrophoresis was first introduced by P. H. O'Farrell and J. Klose in
1975. In the original technique, the first-dimension separation was
performed in carrier ampholyte-containing polyacrylamide gels cast in
narrow tubes.
- The physical
characteristics of proteins are fundamentally important in
organismal function. We used the
complete predicted proteomes of >100 organisms spanning
the three domains of life to investigate the comparative
biology and evolution of proteomes. Theoretical 2D gels were
constructed with axes of protein mass and charge (pI) and
converted to density estimates comparable across all types
and sizes of proteome.
Applications
- Applications of 2-D
electrophoresis include proteome analysis, cell differentiation,
detection of disease markers, monitoring therapies, drug discovery,
cancer research, purity checks, and microscale protein
purification.
- A large and growing
application of 2-D electrophoresis is "proteome analysis." Proteome
analysis is "the analysis of the entire PROTEin complement expressed by
a genOME".
- The analysis involves
the systematic separation, identification, and quantification of many
proteins simultaneously from a single sample. Two-dimensional
electrophoresis is used in this technique due to its unparalleled
ability to separate thousands of proteins simultaneously.
Technology
- 2-D Gel Technology has
had profound impact on proteomic research over the years. Informatics
support brought a new dimension to 2D gels and associated technologies.
But with advent of new and emerging technologies, it will be interesting
to observe the trends of 2D gel technology in the years to come. Here we
review 2D gel technology and its applications besides looking at the
future scope of 2D gels in the post genome era.
- The separation of
proteins based on both molecular charge and size, done by combining the
processes of isoelectric focusing (IEF) and polyacrylamide gel
electrophoresis (PAGE), makes it possible to effectively separate
thousands of proteins using a single gel. This powerful technique,
two-dimensional gel electrophoresis (2DGE) enables scientists to isolate
single or multiple protein targets with a high degree of sensitivity.
- Proteins resolved by
2DE can be identi¢ed based on unique attributes that are measured by MS.
These attributes are determined from analysis of peptides generated by
proteolytic digestion of the protein of interest.
- Proteomics is a field
that is younger than DNA sequencing and although applications are more
limited, the impact of the technology could prove to be enormous. The
industry is growing more by a diffusion process as the technology
gradually replaces older laboratory methods.
Market
- Two-dimensional (2D)
gel electrophoresis can be time-consuming and labour intensive, but its
value to proteomics research means that the market for these products
will more than double by 2010.
- Right now, 2D gel
electrophoresis generates $405.7 million a year worldwide with annual
increases of 12.8%, according to Frost & Sullivan.2 By 2010,
revenues should hit $717.2 million. GE Healthcare (formerly Amersham
Biosciences) and Bio-Rad currently hold 65% of the market.
- Protein microarrays are
a relatively new tool available to scientists. Within the 2005 Protein
Expression and Analysis Survey, we find an equal percentage of
respondents using protein microarrays as are using mass spectrometry in
their protein biomarker research. And given the relative youth of
protein microarrays, a noteworthy percentage (43%) of researchers
surveyed plan to use them this year. It will be interesting to see how
these numbers change and grow in future surveys.
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