The OMRF Multiplexing Protein Quantification Core specializes in targeted-validation proteomics. Targeted-validation proteomics typically follows the discovery phase. For targeted-validation proteomics, specialized mass spectrometers are used to specifically quantify levels of a small set of proteins in a large number of samples. Targeted-validation proteomics generally utilizes 1000s of samples to measure 10s of target proteins.
The OUHSC/OMRF Multiplexing Protein Quantification Core is a fee-for-service core that focuses on targeted-validation proteomics applications. Core Director Dr. Kinter is a nationally recognized leader in the field of targeted-validation proteomics. The core uses two complementary technologies based on LC and either tandem mass spectrometry or high-resolution mass spectrometry based on two different high performance systems: a triple quadrupole mass spectrometer or hybrid quadrupole-Orbitrap mass spectrometer.
In these experiments, the peptides are quantified as chromatographic peaks at characteristic retention times, and the abundance is calculated on the basis of the integrated area of those peaks.
The tandem mass spectrometry experiments are known as selected reaction monitoring (SRM) or parallel reaction monitoring (PRM).
The high-resolution experiments are known as high-resolution accurate mass (HRAM). In each technique, the goal is to specifically detect carefully selected and validated peptides from the protein being measured, formed by tryptic digestion of the complex protein sample, in an LC-MS experiment. The core routinely develops and uses these quantitative assays.
The core has two advanced MS systems used for targeted validation proteomics: (1) a ThermoFisher Scientific TSQ Vantage triple-stage quadrupole mass spectrometer with capillary column HPLC that is equipped with an Eksigent splitless nanoflow HPLC system and ThermoFisher Scientific Proxeon microspray ion sources and (2) a ThermoFisher Scientific Q Exactive Plus hybrid quadrupole–Orbitrap MS system with a Dionex UltiMate 3000 RSLCnano system.