Optimizing All Steps of the Proteomic Workflow for Different Applications

Sample
Collection

  Laser Capture Microdissection of pre-cancerous or invasive cells from matrix tissues. 
Formalin-Fixed Tissue proteomic analysis
HUPO standards for serum and plasma collection

Sample
Preparation

 
Trace Analysis
:  Lysis, digestion, labeling, and LC-MS proteomic analysis of
less than 10,000 cells
Multiple Lectin (MLAC) Chromatography
to extract and fractionate glycoproteins from sera.
Multidimensional Separations using IEF or SCX prior to RPLC-MS analysis.
Tagging of proteins to enhance sensitivity (electrophores, Alexa-fluor)
or to select protein classes of interest (ICAT, iTRAQ,
16O/18O differential labeling). 

   LC and
LC-MS Technologies

 
ERPA
(Extended Range Proteomic Analysis) 95% sequence coverage of large
(180 kDa) proteins, with characterization of phosphorylation and glycosylation. 
14 mass spectrometers, including  LTQ-FTMS and a 2 kHz MALDI instrument constructed in-house. 
Ultranarrow Monolithic Capillary Columns: ultratrace analysis.
Capillary LC and 2D LC  (Advion NanoMate;  LC Packings; Dionex)
NanoSplitter: 1000-fold higher sensitivity in 4 mm LC-MS.

Data Processing
and Analysis

 
MEND
: denoising using information from LC dimension as well as MS dimension. 
PRESEL: optimized selection of precursor ions increases identifications by 30%. 
Linux cluster. 
Additional Capabilities  
Bioorganic chemistry
: immunotoxins; PSA-activated toxins or imaging agents. 
NMR:  ultratrace microcoil NMR, LC-NMR.
Applications  
Biomarker discovery and validation: breast and cervical cancer, cardiovascular disease;   DNA Adducts