Mass Spectrometry and Proteomics Facility
The Mass Spectrometry and Proteomics Core Facility is located in the Cannon Research Center at Carolinas Medical Center. It is a fee-for-service facility dedicated to providing proteomics support to the Atrium Health, University of North Carolina at Charlotte, and other non-profit research organizations in the greater Charlotte region. The goal of the core is to facilitate proteomics analysis of clinical and biological samples by offering state-of-the-art instrumentation and proteomics expertise in preparing, analyzing, and bioinformatic interpretation of experimental data.
The current mass spectrometry instrumentation includes an LTQ-XL and a TSQ Quantum Ultra, both with acquity UPLC systems, and an LTQ-XL Orbitrap-ETD with nano acquity UPLC. Additionally, we have six data analysis workstations.. Software platforms are used for proteomic data analysis include the Bioworks SEQUEST and Proteome Discoverer MASCOT search engines, Sieve and Scaffold Q+S for data compilation and quantification, PEAKS for de-novo sequencing, and the in-house script Intersect for use in comparison of more than two groups of samples. Pathway analysis is carried out using Metacore software.
The Mass Spectrometry and Proteomics Core Facility uses high-throughput nano flow liquid-chromatography tandem mass spectrometry to identify proteins from one dimensional or two dimensional polyacrylamide gel slices. Sample preparation includes electrophoresis of the sample, gel staining and excision of regions of interest. The proteins are then digested within the gel slices using trypsin. Resulting peptides are extracted from the gel matrix and analyzed using reversed phase nano-litter flow liquid chromatography followed by tandem mass spectrometry. Another option for analysis is a technique commonly referred to as MudPIT (Multi-dimensional Protein Identification Technology), which incorporates strong cation exchange as an additional dimension of peptide separation. Spectra are analyzed using the software tools Bioworks, Scaffold, PEAKS for identification of peptides and compilation of experimental data.
Instrumentation:
LTQ ORBITRAP XL ETD:
This instrument combines an ion trap mass spectrometer with collision induced dissociation (CID), higher-energy collisional dissociation (HCD) and electron transfer dissociation (ETD) fragmentation techniques to produce a high resolution instrument with high mass accuracy. This instrument is capable of performing post translational modification analysis, peptide sequencing and identification as well as isotope labeled and label free protein quantification.
TSQ QUANTUM ULTRA:
The Quantum is a triple quadrupole instrument with high sensitivity and a broad mass range. It can be used for highly selective reaction monitoring (H-SRM) experiments for complex sample analysis. It can also be used to study lipids, carbohydrates and nucleic acids, as well as peptides and organic compounds.
ACQUITY AND NANO-ACQUITY UPLC SYSTEMS:
These chromatography systems have the capability of functioning at higher backpressure. They utilize particle chemistry below 2 microns in size. This combination of features enhances efficiency, increases throughput and improves sensitivity for proteomics applications.
Please see our Mass Spec Brochure for more information regarding the use of these instruments in biomedical research.
Laboratory Members
Sunil Hwang, PhD, Director
Kimberly Q. McKinney, MS, Senior Research Analyst
Jin-Gyun Lee, PhD, Post Doctoral Research Fellow
Antonis Pavlopoulos, Research Tech II
Sample Submission and Contact Information
For a fee schedule of services offered by this Core, please click on the following link: Mass Spectroscopy fee schedule. For a template form to accompany sample submission, please click on the following link: Mass Spectrometry Sample submission form.
For more information regarding fee schedule, sample submission, and collaboration, please contact Kimberly McKinney at phone 704-355-5877 or email Kimberly McKinney.