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Interdisciplinary
Materials Research |
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Rapid Protein Mixture Fractionation on RF Plasma Polymer Modified Sample Stages with Analysis by MALDI Mass Spectrometry Dr. Gary Kinsel Now
that the mapping of the genetic sequence of many organisms is well
underway, many bioanalytical laboratories today are involved in the
daunting task of attempting to characterize all proteins expressed by a
given biological systems i.e., the so-called cellular proteome. In
short, it is believed that mapping of targeted cellular proteomes,
under various conditions, has the potential to yield new approaches for
the diagnosis of disease, new targets for drug therapy, and new
detection tools for chemical/biological weapons, among a host of other
important outcomes. In these proteomic characterization efforts
biological mass spectrometry (MS), and in particular matrix-assisted
laser desorption / ionization (MALDI) MS, has emerged as an essential
tool, allowing characterization of large numbers of proteins directly
extracted from various organisms.
The
research in our group focuses on expanding the utility of MALDI MS for
these proteomic investigations through the development of
high-performance surface modified MALDI probes that allow the rapid
on-probe fractionation and analysis of the complex mixture of proteins
targeted in a typical investigation. Our approach to the development of
these devices involves the use of a radio frequency plasma to deposit
polymer thin films directly on the surface of MALDI probes and the
optimization of the conditions used to chemically or
 bioselectively
fractionate protein mixtures prior to MALDI MS analysis. The rf plasma
polymer deposition approach allows us to explore the utility of an
extraordinarily rich array of surface chemistries for these proteomic
fractionation applications, including non-fouling surfaces, temperature
responsive surfaces, solvent responsive surfaces, etc. Our results
confirm that this on-MALDI-probe fractionation approach to the
characterization of cellular proteomes provides substantial
enhancements in the information content of the mass spectral data and
can be successfully used for the discovery of biomarkers indicative of
cellular stress, disease, etc. REU
students will find themselves in an extraordinarily rich research
environment, with the opportunity to gain hands-on experience in any of
a number of areas, including modern mass spectrometry instrumentation,
RF plasma polymer deposition equipment, thin-film surface
characterization tools and cutting edge biological analysis apparatus.
Specific projects that will involve REU students include (1)
optimization of on-MALDI-probe fractionation conditions using control
mixtures of peptides and protein, (2) isolation and mass spectrometric
identification of bacterial peptides/proteins showing distinctive
fractionation behavior within behavior/structure correlation studies,
and (3) creation and characterization of new RF plasma polymer surface
films. These areas of research are well-established and the REU student
will find themselves in a position to make contributions of
significance to a variety of future professional publications and
presentations.
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