Isotope coded protein label technology differs from other approaches in that the labelling step is not limited to the peptide level but was developed to be applied on the protein level
Isotope coded protein label (ICPL) technology enables true quantitation of differentially expressed proteins with an unmatched dynamic range due to the optional combination with intact protein fractionation steps.
In contrast to other isotope labeling approaches, the ICPL labelling step is not limited to the peptide level but was developed to be applied on the protein level.
For complex proteomes, the complexity of the resulting peptide mixtures typically exceeds the capacity of 1D or 2D peptide chromatographic steps and requires reduction of complexity.
As a unique feature, ICPL allows such reduction of complexity on the protein level by chromatographic or other fractionation steps without any sacrifice on quantitation accuracy.
All lysine side chains are modified by the ICPL tag, introducing a mass difference of 6Da per labelled site.
Multiple peptides from each protein usually carry the ICPL- tags after proteolysis which allows robust statistics.
The ICPL chemistry even leads to the detection of modified peptides with increased sensitivity in MS.
The ICPL workflow is compatible with all established LC- or gel-based protein and peptide fractionation and separation techniques, and with all mass spectrometer types on the market including ion traps.
Protein identification and quantitation can be obtained after protein digestion either with ESI-MS/MS on Bruker Daltonics HCTultra ion trap or Microtof-q Q-q-TOF system, or using Maldi-MS/MS on the Ultraflex II Tof/Tof.
ICPL-based quantitative analysis is fully supported by Bruker Daltonics's Proteineer-LC 1.1 suite providing automated analysis and data validation.
Through the co-marketing agreement between Bruker Daltonics and Serva, the Serva ICPL kit can now be obtained worldwide from Bruker Daltonics.
Detlev Suckau, head of proteomics development at Bruker Daltonics, stated: "We consider the new ICPL labelling technology a breakthrough for true quantitation in proteomics and biomarker discovery.
"For the first time, entire proteomes can be analysed quantitatively and on solid statistical grounds.
"The bioinformatics packages within our Proteineer-LC 1.1 suite will help our customers to run ICPL analyses very conveniently in order to obtain validated data quickly.
"The ICPL kits represent a great step forward towards high-content proteomics in combination with our outstanding LC-MS/MS instrumentation.
"We are very pleased and look forward to a productive collaboration with Serva Electrophoresis".
Barbara Mueller of Serva Electrophoresis said: "Serva is a leading supplier of electrophoresis specialty products, and is the exclusively licensed distributor of the unique ICPL kits.
"Under the co-marketing agreement with Bruker Daltonics, new avenues of application are expected with a clear benefit to the proteomics community".
ICPL inventor Professor Friedrich Lottspeich commented: "Key to the success of proteomics technology is that quantitative information on proteins can be obtained fast and reliably with a high dynamic range in order to access low abundant proteins.
"These are typically the proteins that matter.
"That absolutely requires the proteome fractionation on the protein level and not only on the peptide level.
"The ICPL technology was designed to provide reliable quantitation due to data redundancy and the compatibility with any combination of protein fractionation technologies, including 1D or 2D gel electrophoresis or protein LC.
"I am delighted that this powerful technology will now be made available to the scientific community"
