Capillary-Channeled Polymer (C-CP) Films as Processing Platforms for Protein Analysis by Matrix-Assisted Laser/Desorption Ionization Mass Spectrometry (MALDI-MS) |
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Authors: | Jennifer J Pittman Benjamin T Manard Paul J Kowalski R Kenneth Marcus |
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Institution: | (1) Department of Chemistry, Biosystems Research Complex, Clemson University, Clemson, SC 29634, USA;(2) Bruker Daltonics, Inc., Billerica, MA, USA; |
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Abstract: | Polypropylene (PP) capillary-channeled polymer (C-CP) films have parallel, μm-sized channels that induce solution wicking
via capillary action. Efficient mass transport from the solution phase to the channel surface leads to adsorption of hydrophobic
protein solutes. The basic premise by which C-CP films can be used as media to manipulate analyte solutions (e.g., proteins
in buffer), for the purpose of desalting or chromatographic separation prior to MALDI-MS analysis is presented here. Cytochrome
c and myoglobin prepared in a Tris-HCl buffer, and ribonuclease A, lysozyme, and transferrin prepared in phosphate buffered
saline (PBS), are used as the test solutions to demonstrate the desalting concept. Protein analysis is performed after deposition
on a C-CP film with and without a water washing step, followed by spray deposition of a typical sinapinic acid matrix. Extracted
MALDI mass spectra exhibit much improved signal-to-noise characteristics after water washing. A mixture of cytochrome c and myoglobin (2 μL of 2.5 μM each in Tris-HCl buffer) was applied, washed with water and spatially separated via simple
capillary action (wicking) using a reversed-phase solvent composition of 0.1% trifluoroacetic acid (TFA) in 50:50 acetonitrile
(ACN):H2O. Subsequent application of sinapinic acid followed by imaging of the film using MALDI-MS reveals that as the protein solution
is wicked down the film, separation occurs. |
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