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Information @ a Glance

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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