NEW YORK, April 17, 2015 /PRNewswire/ -- Advanced capillary electrophoresis (CE) systems that can define thousands of proteins at once and separate them quickly are being put to use for the new methods of drug development, one where proteins are analyzed intact. According to Kalorama Information, the methods of biomarker development are changing, and with it, a possible change in the preferred technology. The firm has studied mass spectrometry markets several times over the past few years and noticed the preference of the use of CE over liquid chromatography methods in research for an upcoming proteomics discovery report.
The bottom-up shotgun approach has powered over a decade of extensive research in proteome mapping and protein discovery for biomarker and drug development. However, bottom-up approaches do not lend themselves well to the differentiation and discovery of proteoforms, or the different in vivo forms the same protein. Top-down proteomics or the analysis of intact proteins is a less mature approach, but could contribute significantly to systems biology understanding and the characterization of drug targets in the drug discovery process.
Proteomics Markets for Research and IVD Applications, Kalorama's latest reporting on proteomics, can be found at: http://www.kaloramainformation.com/redirect.asp?progid=87335&productid=8359865.
"Much as robust bottom-up proteomics came about through the successful interface of nanoscale LC and electrospray, the future of top-down proteomics may hinge on the demonstration and standardization of CE-electrospray ionization (ESI)-MS systems," said Emil Salazar, Kalorama analyst and author of an upcoming report on Discovery Proteomics, a recent blog post on Kalorama Information.com "One obstacle to higher efficiency top-down proteomics studies is effective sample separation, which has motivated recent research into capillary electrophoresis (CE) methods."
There are several factors make CE, particularly capillary zone electrophoresis (CZE), attractive or superior to LC for use in top-down proteomics:
- CE mitigates weak bonding or analyte interaction with separation media as in the case with LC columns; interaction between proteins or polypeptides and LC separation media impacts sample recovery and separation efficiency
- Better handles large proteins and polypeptides (+50 kDa); large proteins in reverse-phase LC (RPLC) columns can take longer and are more difficult to elute, produce broad spectrum peaks, and reduce column lifetime
- Faster separation can be accomplished using CZE than HPLC or within 10-30 minutes for most elutions compared to 50 minutes with HPLC
- Greater sensitivity with resolution of analytes in the low nanogram range compared to the microgram-high nanogram range of HPLC
- Higher efficiency separation is possible with theoretical plate counts approaching 100,000
- Utilizes very small amounts of sample with CZE loading capacity one to three orders of magnitude lower than HPLC
Several research programs are focused on optimizing CE technology and analytical practices for top-down proteomics. Much of the published research on top-down proteomics using CE has been in the last three years from researchers at institutes including Notre Dame , Northwestern, and at the Scripps Research Institute. A solution to the minute sample loading capacity of CZE is coupling the separation method with capillary isoelectric focusing (CIEF) or another second dimension of separation, though at a cost of added complexity in sample manipulations. Work also continues on optimizing CZE, as the simplest, most common form of capillary electrophoresis, for top-down proteomics. Importantly, researchers have been able to apply and refine electrospray ionization (ESI) with a sheathless design for on-line CZE-ESI-MS development.
An upcoming title from Kalorama Information will explore the markets for proteomics research instrumentation with a thorough accounting of the capabilities, specific applications, and development of technologies such as capillary electrophoresis, mass spectrometers, and liquid chromatography. Proteomics Markets for Research and IVD Applications (Mass Spectrometry, Chromatography, Microarrays, Electrophoresis, Immunoassays, Other Technologies), which is a broader market outlook for proteomics including clinical IVD products, is available with http://www.kaloramainformation.com/redirect.asp?progid=87335&productid=8359865.
About Kalorama Information
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SOURCE Kalorama Information