Large-scale study of proteins, as a growing field in biological sciences, has gained much attraction in recent years. One of the most important factors in success of the field is the evolution of novel techniques for the separation and identification of proteins. The most extensively used techniques in biochemistry and molecular biology is electrophoresis, with gel electrophoresis (GE) and capillary electrophoresis (CE) being the most preferred choice of researchers for efficient separation of macromolecules.
There have been considerable technology advances in electrophoresis although it is a standardized technique because of its ever-expanding research applications. Among the major GE techniques, the polyacrylamide gel (2-D PAGE) technique has received immense attention due to its high resolving power to separate posttranslational protein entities. Technology enhancement methods like 2-D fluorescence difference gel electrophoresis (2-D DIGE) hold promising protein detection.
There have been significant technology developments in informatics and gel imaging. Technology developments in gel imaging are accompanied by corresponding progress seen in gel informatics. Informatics vendors are taking many initiatives to address the growing concern of reproducibility in the 2-D gel technique by sophisticated signal-analysis algorithms that reflect true biological outcomes. Gel informatics is likely to witness highest growth compared to other products in the coming years.
The goal is to perform electrophoresis with minimal resources - primarily in terms of time and labor without impacting reliability or reproducibility. The need for quality and improved efficiency is driving all purchasing decisions and are extending the application scope of electrophoresis to new areas.
As time progresses, the wide selection of products and tools available that help laboratory bench workers perform electrophoresis continue to become more refined and improved.
Expanding upon the innovative first-generation iBind Western System, Thermo Fisher Scientific has developed the new iBind Flex Western System for automated western-blot processing. Based on proprietary sequential lateral flow, the latest platform is designed to enable a more versatile walk-away solution for the immune-detection step in a western-blotting workflow. Researchers now have the ability to adapt the iBindTM Flex Western device to a variety of blot formats and sample throughput. Compatible with downstream chemiluminescent, colorimetric, or fluorescence-detection protocols and optimized for higher sensitivity and reproducibility, the device can reduce the amount of primary antibody required when compared to manual blot processing.
Another new offering is by Cleaver Scientific, a horizontal polyacrylamide gel electrophoresis (hPAGE) system. While vertical systems are commonly used for polyacrylamide gel electrophoresis, this unit's horizontal format allows for a large number of samples to be run simultaneously and also offers greater resolution through use of polyacrylamide instead of agarose. The system's main advantage is the simplicity of pouring and casting the polyacrylamide gels. This takes away some of the complexity normally associated with vertical gel casting.
For researchers who need to separate samples with very similar molecular weights, the new Clarit-E Choice Stretch electrophoresis gel tanks from Alpha Laboratories Ltd. provide the perfect solution. Plus, when high throughput is needed, these tanks allow up to 350 samples to be resolved per gel. The extended run lengths of the gel tanks allow restriction fragments or other sample bands of comparable molecular weight to be easily separated and identified, making them ideal for researchers wanting to perform higher-resolution separation of more samples over a longer distance. Speed-loading is accomplished using 10, 14, 16, 18, 28, or 30 sample multi-channel pipette-compatible combs.
The traditional hazards of DNA gel electrophoresis lie in the use of ethidium bromide stain, a known mutagen and carcinogen, and of UV illumination sources, which can cause eye and skin damage.
In all capillary-based electromigration techniques, including capillary gel electrophoresis, capillary isotachophoresis, capillary isoelectric focusing, micellar electrokinetic chromatography, affinity capillary electrophoresis as well as in the hybrid techniques progress has been made in experimental setups, and for many groups of analytes, such as peptides, proteins, nucleotides, saccharides, drugs and their metabolites, and CE/ESI-MS, it has been successfully applied. Electromigration is also further miniaturized. New pre-concentration methods are allowing the investigation of compounds, which are not sensitively detected with ESI-MS. Coordination ion spray (CIS) MS is another method for observing growth of sensitivity-enhancement by online formation of charged coordination compounds.
Challenges and Opportunities
Rapidly emerging economies, rising demand for personalized medicine, rising funding for research on electrophoresis techniques, growing number of research collaborations between market players and academic institutions, increasing use of CE with mass spectroscopy, growing focus on next-generation sequencing research, and a shift from plant-derived to genome-based drugs has created a buzz in the industry, thereby creating an array of growth opportunities.
2-D electrophoresis is widely applied and remains the method of choice in proteomics; however, pervasive 2-D electrophoresis-related concerns undermine its prospects as a dominant separation technique in proteome research. Consequently, the state-of-the-art shotgun techniques are slowly taking over and utilizing the rapid expansion and advancement of mass spectrometry to provide a new toolbox of gel-free quantitative techniques.
Several alternatives to electrophoresis technologies are available in the market, which offer better efficiency and results. In addition, some electrophoresis techniques involve time-consuming operations and offer limited sample analysis, limitations on sample analysis through gel electrophoresis, and safety concerns related to mutagenic nucleic acid dyes are some of the restricting factors affecting the growth of electrophoresis market.
In the post-genomic era, when several proteomes are on the verge of completion, the promising field of protein-based diagnostic techniques is emerging. Although protein detection has been used for a long time in clinical diagnostic tests; yet high-throughput proteomics approaches along with systems biology could be a step forward, toward the development of next-generation diagnostic tools and pave the way for personalized medicine.
Even after three decades of development and modification, the comet assay is still a rather simple, versatile, but labor-intensive assay. Both in vivo and in vitro applications could gain great advantage from further improvements in efficiency, standardization of protocol, and throughput. Acceptance of the in vitro comet assay for genotoxicity testing, inexpensive automated comet scoring to save researchers from interminable microscope viewing, protocol standardization and reliable internal reference standards, more human bio-monitoring studies of DNA repair, environmental monitoring using a variety of animal and plant species, and many more unpredictable developments and applications might be observed in the next couple of years.
Looking at recent researches and products entering the market, lab-on-a-chip might change the way for diagnostics in the near future. The challenge for industrial research is to incorporate on the same lab-on-a-chip the maximum amount of individual operations in order to decrease costs and increase ergonomics and the speed of diagnosis. At the moment, technologies are not unified and nobody can say which technologies and which materials will be the most promising for high-throughput diagnostics. Therefore, in future, manufacturers may provide instruments that are efficient, accurate, and competitively priced to increase productivity of laboratories and thereby encourag the analyzer demand.