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A bright future ahead for separation science

With high versatility, major advancements have been made in the instrumental set-ups over the years. The changes in technology trends of electrophoresis suggest that the next step would be miniaturization and portability of the systems.

Electrophoresis was introduced in the beginning of the nineteenth century, and even after the passing of two centuries, it remains as relevant as ever. Although electrophoresis is presently being done in many different ways and methods that the equipment and style is quite different from the original design, the core principle remains the same. The changes in technology trends suggest that the next step would be miniaturization and portability of the systems.

Global market
The electrophoresis market is expected to reach USD 3.6 billion by 2025 from USD 2.7 billion in 2020, at a CAGR of 6 percent, estimates MarketsandMarkets. The rising incidence of cancer, infectious diseases, and genetic disorders, growth in funding for research on genomic, proteomic, and electrophoresis techniques, the growing number of industry-academia research collaborations, the growing use of capillary electrophoresis with mass spectroscopy, the increasing use of next-generation sequencing, and the rise in the number of clinical, forensic, and research laboratories are factors driving the growth of the electrophoresis market.

The reagents segment accounted for the largest share of the market in 2019, by product. This can be attributed to the increasing use of consumables with the rise in electrophoresis in the fields of proteomics, genomics, drug discovery, antibody development, and personalized medicine, among others. Also, the increasing demand for 2D electrophoresis for protein separation in a number of applications such as drug discovery, protein mapping, and the diagnosis of chronic diseases, along with the increasing application areas of capillary electrophoresis are supporting the market growth.

The research segment accounted for the largest share of the market in 2019, by application. The large share of this segment can be attributed to the increasing application of electrophoresis in the field of drug discovery, proteomics, genomics, and antibody research. Moreover, increasing research in the field of biomarker discovery and NGS is increasing the adoption of electrophoresis systems and consumables.

With increasing research being conducted in the fields of drug designing, proteomics, genomics, sequencing, biomarker discovery, and personalized medicine, the demand for electrophoresis products is expected to increase in academic and research institutes.

The electrophoresis market in the Asia-Pacific is expected to grow at the highest CAGR during 2020–2025. This region is expected to grow at the highest pace during the forecast period, primarily with growing proteomics and genomics research, increasing investments by pharmaceutical and biotechnology companies, growing awareness about personalized therapeutics, and increasing research activities in the field of mAbs-based therapeutics.

Prominent players in the global electrophoresis market are Thermo Fisher Scientific (US), Bio-Rad Laboratories (US), Merck Group (Germany), Agilent Technologies (US), Danaher Corporation (US), GE Healthcare (US), PerkinElmer (US), Qiagen N.V. (Netherlands), Harvard Bioscience (US), Shimadzu Corporation (Japan), Lonza Group (Switzerland), Sebia Group (UK), C.B.S. Scientific Company (US), Helena Laboratories (US), Takara Bio (Japan), Syngene (UK), Teledyne Technologies (US), VWR International (US), Analytik Jena (Germany), and TBG Diagnostics Ltd. (Australia).

Technology trends
With the high versatility, major advancements have been made with regard to the instrumental set-ups over the years. The focus of the industry remains to come up with the best technologies that offer high throughput and improved sensitivity. New strategies have been proposed to develop high-sensitive, portable, or miniaturized devices.

Affinity electrophoresis. Affinity electrophoresis system is a technique in which the resolving capability of capillary electrophoresis is used to separate samples that undergo specific or non-specific affinity interactions during the process of electrophoresis. This process can occur in either solution or immobilized to a solid support. It is also used to measure the binding affinity of receptors to neutral and charged ligands. The use of affinity electrophoresis is extensive in that it is able to detect affinity interactions in either free or immobilized form. Some of its uses include detection for peptides and proteins, drug development, detection of small molecules, and also for immuno-affinity works. There are also different types of affinity systems. Some of the advantages of using affinity electrophoresis is that the sensitivity of the technique allows for more precise detection and discrimination of normal and carcinogenic proteins from the same sample. Also, its wide field of usage makes it an important technique for sampling and data collection.

Automated electrophoresis system. With the advancement of technology, it is now possible to conduct electrophoresis by using computerized robotics and programming that enables electrophoresis protocols to be conducted automatically. Automated systems come in many different types and forms. With today’s technological advances, various types of electrophoresis systems can be automated. Automated electrophoresis systems are also highly accurate and precise and can even detect single-strand conformation polymorphism in genetic samples. One of the new automated systems is the automated buffer-less electrophoresis system, which uses pre-casted gel, either SDS or agarose, that does not require any buffer to run, which is one of its biggest advantages. The system boasts the possibility to view the progress of the gel run by attaching the machine to an ultra-violet light or blue-light viewing add-on. It is capable of separating different types of samples by using the appropriate gel type at high speed.

Microchip electrophoresis. A further advancement to the capillary electrophoresis system, the microchip electrophoresis system boasts a more efficient system. One of the main benefits is an increase in throughput by many folds over the capillary electrophoresis system as the microchip system contains numerous microchannels, which allow high-throughput experiments to be conducted quickly and efficiently. Another benefit is the low fabrication cost as the intricate enclose microchannel is comprised of glass or fused silica substrates. These materials enable ultra-fast DNA separations as the sample-loading format is unique, coupled with short separation distances and optimal thermal characteristics of the glass or fused silica substrates. The microchip electrophoresis system is also fully automated, from sample handling to data analysis, which allows minimal human handling and possible error. Example of this system is the lab-on-a-chip device. Microchip electrophoresis uses laser-induced fluorescence and electrochemical detection method as their detection method. This is due to their extremely small size, and the accuracy needed to read their results require highly accurate methods for microchip electrophoresis to work. Research in developing new materials for microchip electrophoresis is widely delved into by many groups of researchers. Many different types of materials have been used to fabricate the microchip, from the most common material like silica and glass substrates, to poly (dimethylsiloxane), or PDMS, and poly (methylmethacrylate), or PMMA, as a material to fabricate microchip electrophoresis via thin-casting method. The advantage of this protocol reduces the cost of production, and also allows faster results development.

FACE. Fluorophore-assisted carbohydrate electrophoresis (FACE) is used to identify carbohydrates with an attached fluorescent dye by separating the carbohydrates, using a polyacrylamide gel. This technique is important as carbohydrates are not charged, and it is the main technique used to analyze different types of carbohydrates like glycoproteins, glycolipids, plant, and bacterial polysaccharide.

CE-MS. The last decade has witnessed a revival in technological developments for CE-MS, while at the same time moving away from being a purely academic tool to one increasingly being adopted by industrial scientists. Until a few years ago, there was only one commercial supplier offering a technology solution, meaning little progress. But more recently, there have been new players entering the market with new technologies. This will get better and better, making CE-MS more sensitive and more robust. Some players are making impressive strides toward miniaturizing the technology with the development of microfluidic CE-MS. Miniaturization will enable a lot of high-throughput work, without compromising the level of information they offer. This means that any process one wants to monitor will become more cost-effective.

High-speed CE with LIF detection. Currently, the miniaturization of high-speed CE systems has become one of the major development directions of technology, which can provide various portable instruments for point-of-care testing, in-situ analysis, and extraterrestrial exploration by means of their small size, high-resolution separation, and fast analysis time. So far, most of the miniaturized high-speed CE systems are developed on the basis of microchip-based CE technique with advantages of automated picoliter-scale sample injection and separation and high system integration. In the early stage of the development of miniaturized CE instruments, electrochemical detectors were adopted frequently owing to their simple structure and small size.

Buyers’ perspective
Lab managers evaluate instrumentation based on its capabilities, not its buzz factor. Purchasers of chip-based systems should, therefore, make sure that systems under consideration provide the depth and breadth of assays they expect to run.

Systems that offer assays appropriate for sample type, especially with regards to sample concentration, sample size, sample type (DNA, RNA, protein, etc.) are recommended. Also, consider the vendor’s services for support, warranty, repair, and installation, and the system’s ease of use, all of which can help reduce lab downtime to a minimum.

Sample volumes have become a major selling point with all analytical instruments. Smaller volumes consume less reagent and less sample, and allow researchers to do more with rare or scarce samples. Some systems, for example, require just one microliter of sample for RNA and DNA assays.

Regulatory compliance is an issue for pharmaceutical R&D labs, with 21 CFR part 11 being the applicable regulation covering electronic records. Compliance also plays into standard operations for diagnostics labs and any facility with ongoing interaction with the patent or legal system, for example, forensics.

Cost is always a consideration, but for instrumentation that labs use every day, cost of ownership must be considered with regard to energy and reagent consumption. Service plans tend to be comprehensive, with generally rapid response, but very busy labs should also consider the economic impact of downtime.

Way forward
With the development of increased sensitivity in detection systems, it is possible to increase the speed and processing of electrophoresis. This also enables researchers to use less of their samples, which is the best advantage of capillary electrophoresis, as it will mean less preparation time and also less wastage of sample.

Although the technology of electrophoresis has advanced tremendously from the basic paper electrophoresis system to today’s highly advance microchip electrophoresis system, there is still a basic need for an external power supply to run the electrophoresis, more so as the system becomes more sophisticated. This serves as a bottle neck on the usage of any electrophoresis system in that the usage of any system is limited to a room with an external power supply. Without a power supply, electrophoresis cannot be conducted. Active research is being carried out currently to overcome these limitations.

Other disadvantages of the current electrophoresis systems and techniques include the influence of contaminations such as vanadium and selenium can compromise gel results, especially if the process is left to run for a long period of time. However, more testing is needed to be carried out regarding the effect of contaminations on gel results’ viability.

With capillary electrophoresis, a lot of care into the details, such as capillary position and gas flow rates, of each experiment runs have to be taken to be able to reproduce the same results as differences in the settings will generate different results for the same experiment.

With advancement in technology, both in engineering and sciences, it is now possible to produce smaller circuit boards that are light and have higher efficiencies and more efficient transformers. Portable electrophoresis system could be built on these. Currently, there is preliminary research into designing such a system, but the efficiency of such systems is still being tested.

Also, there are many patents regarding increasing the efficiencies of current systems, such as using a different buffer system or gel type, in increasing the efficiency. There is major research being done by different groups around the world on improving currently used systems, notably the automated electrophoresis system. The main point of the research being done is to increase the detection limit and also the reduction in the amount of background noise. Also, the uses of different types of materials, like coated capillaries, for running gel electrophoresis and upgrading the detection system on the current equipment are also being looked into. The materials being researched and developed by these groups mainly focus on increasing the efficiencies, reproducibility, and accuracy of the separation run. Work is also being done to enable retrieval of the separated sample without damaging it.

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