Indiana Section of the Society for Applied SpectroscopyIndiana Section of the Society for Applied Spectroscopy
2001-2002 Seminar Abstracts
A 2-D LIQUID SEPARATIONS/MASS MAPPING METHOD FOR INTERLYSATE COMPARISONS OF CANCER PROGRESSION
Dr. David M. Lubman
Department of Chemistry, The University of Michigan
Ann Arbor, Michigan 48109-1055
http://www.geocities.com/ResearchTriangle/Facility/2190/
A
novel 2-D liquid separations/mass mapping method is being developed to profile
the protein content of tumor cells. The method involves using 2-D liquid
separations as a replacement for 2-D gel electrophoresis, where the first
dimension involves a liquid phase batch IEF separation of proteins according to
their pI and the second dimension is based on separation by nonporous
reversed-phase HPLC. The result is a 2-D comprehensive separation of proteins
where the proteins can be collected as isolated proteins in the liquid phase as
the eluent of HPLC. Alternatively, the liquid eluent can be detected on-line
using electrospray TOF mass spectrometry to provide the molecular weight of the
intact proteins. The result is a 2-D mass map based upon pI versus MW, which is
analogous to a 2-D gel but which provides MW accurate to a 100 ppm accuracy. The
advantages of this method in terms of speed, throughput, sensitivity, and
quantitation will be discussed. Further
it will be shown that by using these highly accurate mass maps that interlysate
comparisons can be performed to search for changes in protein expression and
structure as a function of the progression of cancer.
Applications to protein profiling for biomarkers related to the detection
and prognosis of ovarian and breast cancers will be discussed.
Contact Indiana Section of the Society for Applied Spectroscopy