7:45 Morning Coffee or Breakfast Workshop (Sponsorship Available)
8:25 Chairperson’s Remarks
8:30 Glycans in Hepatocellular Carcinoma
Radoslav Goldman, M.D., Assistant Professor, Oncology,
Georgetown University Our glycomic study of hepatocellular carcinoma (HCC)identified novel candidate markers for early detection of the disease.
Glycans enzymatically released from serum proteins were analyzed by MALDI-TOF/TOF following solid phase
permethylation. Analysis of less than 0.05
ml of serum allowed relative quantification of 80 glycans. A set of 3 glycans is
sufficient to classify HCC with 93 % prediction accuracy in a blinded validation
set of samples (n=75). In conclusion, we identified a set of novel candidate
markers for early detection of HCC. The results provide leads for further
improvement of analytical strategies in biomarker discovery.
9:00 Biomarker Proteomics: Pitfalls and Potentials
Niels Heegaard, Professor, Director, Autoimmunology, Statens Serum Institut Biological fluids contain a largely uncharacterized reservoir of biological
information through their content of a multitude of proteins and peptides, i.e.
their proteomes. MS-based proteomics has a digital output and promises to
accelerate the discovery of diagnostic biomarkers through the ability to measure
many analytes in one operation. However, such analyses require strictly
controlled clinical designs, sampling, sample handling, and data processing,
i.e. variable selection (peak picking). We illustrate these issues and results
from a large study of sera from patients suspected of ovarian cancer.
9:30 Straightforward Strategies for Reducing Sample Complexity
Janice Simler, Ph.D., Research Scientist III, Bioscience, Millipore We propose a strategy for reducing sample complexity using
ultrafiltration and affinity based scalable depletion. Serial enrichment using
ultrafiltration centrifugal devices with decreasing molecular weight cutoffs
allows the study of particular ranges of the proteome. Depletion of several
selectable high abundance proteins enables the study not only of the lower
abundance proteins but also the low abundance proteins that may be
non-specifically interacting with high abundance proteins, such as albumin.
Combining serial ultrafiltration with the affinity based scaleable depletion can
further enhance sample complexity reduction.
10:00 Solution Showcase
(Sponsorship
Available)
10:15 Coffee Break in the Exhibit Hall
Tissue Proteomics
11:00 Comprehensive Analysis of White Fat Adipose Tissue Using
Detergent-Free Protein Extraction by Pressure Cycling and High Resolution
Tandem Mass Spectrometry Alexander Ivanov, Dr., Research Scientist, Director, Harvard NIEHS
Center for Environmental Health Proteomics Facility, Harvard School of Public
Health Fat adipose tissue plays a key role in energy metabolism,
lipid synthesis and secretion of signaling proteins linked to obesity, insulin
resistance, inflammation and other physiological complications. Efficient
proteomic analysis of adipose tissue is highly valuable for studies of these
diseases. Fat adipose tissue contains up to 80-90% lipids, which makes
conventional detergent-based protein solubilization and extraction methods
inefficient. This study was enabled by the use of alternating hydrostatic
pressure and specialized organic solvents for disruption of cells, micelles and
membrane fragments and efficient protein recovery from lipid-rich adipose tissue
followed by 1D- and 2D-SDS-PAGE and protein identification by liquid
chromatography and high performance tandem mass spectrometry.
11:30 Unlocking Tissue Proteomics for Biomarker Discovery Cheng S. Lee, Ph.D., Chief Executive Officer, Calibrant Biosystems;
Associate Professor, Department of Chemistry and Biochemistry,
University of Maryland There is increasing acceptance that relating tissue
morphology with data obtained from various molecular analyses is critically
important. Furthermore, proteomic profiling of archived formalin-fixed tissues
for which there is corresponding clinical information has great potential for
biomarker identification. Such biomarkers may have accurate, consistent,
precise, and reproducible diagnostic, prognostic, or therapeutic utility in
clinical settings. The GeminiTM technology is a powerful proteome
platform to investigate the effects of fixation and archival time on subsequent
antigen retrieval. In addition, tissue proteome analysis can be conducted using
archival tumor tissue blocks prepared as far back as the early 1980s.
12:00 pm LCM, the Necessary Technology for Individualized Therapy
Amy VanMeter, Research Specialist, Center for Applied Proteomics and
Molecular Medicine, George Mason University Laser Capture Microdissection (LCM) is a powerful tool for
procuring purified cell populations from heterogeneous tissue under direct
microscopic visualization. This technology is essential for molecular profiling,
compared to direct extraction of the tissue, because the diseased cells of
interest (e.g. cancer cells) in the tissue constitute a small (or unknown)
proportion of the tissue sample being analyzed. The presentation will begin with
an overview of the latest technology in Laser Microdissection, with an emphasis
on the downstream analysis of molecules procured by this technology. Combining
LCM with proteomic analysis offers an exciting approach to map the activity of
kinase signaling pathways which are the protein drug targets as the basis for
individualized therapy. In this regard, example studies will be presented
describing signal pathway mapping of Non Small Cell Lung Cancer and core needle
biopsies of breast cancer.
