This continuation proposal aims at the design, preparation, and testing of completely new monolithic columns in capillary and thin layer formats that will provide unmatched potential for the study, isolation, separation, detection, and identification of biologically active molecules and enable to fully exploit the rich potential of studies in the life sciences. The new monolithic columns and layers will be prepared via a simple free radical polymerization process initiated both thermally and photochemically. One of our main targets will be design and development of macroporous monolithic column suitable for the separation of small molecules and stable in the entire pH range using a sacrificial scaffold approach. Another target will involve novel narrow bore open tubular capillary columns with inner walls covered with a thin layer of porous monolithic polymer. The pore surface of the capillary monolithic columns will then be used directly for high efficiency separations, or modified through UV initiated photografting to incorporate a wide variety of high-density, tailor-made functionalities including hydrophobic, ionizable, and reactive groups. These capillary columns will be specifically designed to achieve the very high efficiency separations of difficult protein and peptide mixtures in nano LC mode. Finally, new approaches to thin layer chromatography and related methods making use of macroporous polymer thin layers will focus on new, extremely thin layers for the 1-D and 2-D separations of biomacromolecules such as peptides and proteins and their direct detection using MALDI-TOF MS. The mobile phase will be driven by capillary forces, pressure, and/or electroosmotic flow. In order to achieve MALDI detection of midsize and large molecules without the need for a low molecular weight matrix, this proposal also targets the development of thin layers including specifically designed monomers carrying functionalities enhancing desorption and ionization of biological samples. These monolithic plates will be prepared using both copolymerization and photografting techniques. Direct use of these plates in mass spectrometer will simplify and accelerate proteomic research. Overall, our targets are to extend the applications of monolithic materials providing them with both enhanced performance and unexpected capabilities in new applications and to demonstrate their vast potential in various areas. This proposed research focuses on the development of monolithic polymers that will serve as separation media, adsorbents, and supports providing real benefits to both the scientists who perform ever more demanding separations with ever smaller amounts of complex samples as well as the engineers who design advanced separation units. Our results can also be used for the engineering of new separation and detection devices suitable for separation of very complex samples such as proteomes, in biochemical assays, and in early diagnostics of diseases with direct impact on affordable health care.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM048364-18
Application #
7753862
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Edmonds, Charles G
Project Start
1993-01-01
Project End
2011-12-31
Budget Start
2010-01-01
Budget End
2010-12-31
Support Year
18
Fiscal Year
2010
Total Cost
$307,282
Indirect Cost
Name
University of California Berkeley
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
124726725
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Lv, Yongqin; Lin, Zhixing; Tan, Tianwei et al. (2014) Preparation of reusable bioreactors using reversible immobilization of enzyme on monolithic porous polymer support with attached gold nanoparticles. Biotechnol Bioeng 111:50-8
Svec, Frantisek (2012) Quest for organic polymer-based monolithic columns affording enhanced efficiency in high performance liquid chromatography separations of small molecules in isocratic mode. J Chromatogr A 1228:250-62
Lv, Yongqin; Alejandro, Fernando Maya; Fr├ęchet, Jean M J et al. (2012) Preparation of porous polymer monoliths featuring enhanced surface coverage with gold nanoparticles. J Chromatogr A 1261:121-8
Urban, Jiri; Svec, Frantisek; Frechet, Jean M J (2012) A monolithic lipase reactor for biodiesel production by transesterification of triacylglycerides into fatty acid methyl esters. Biotechnol Bioeng 109:371-80
Lv, Yongqin; Lin, Zhixing; Svec, Frantisek (2012) Hypercrosslinked large surface area porous polymer monoliths for hydrophilic interaction liquid chromatography of small molecules featuring zwitterionic functionalities attached to gold nanoparticles held in layered structure. Anal Chem 84:8457-60
Lv, Yongqin; Lin, Zhixing; Svec, Frantisek (2012) ""Thiol-ene"" click chemistry: a facile and versatile route for the functionalization of porous polymer monoliths. Analyst 137:4114-8
Teisseyre, Thomas Z; Urban, Jiri; Halpern-Manners, Nicholas W et al. (2011) Remotely detected NMR for the characterization of flow and fast chromatographic separations using organic polymer monoliths. Anal Chem 83:6004-10
Chambers, Stuart D; Svec, Frantisek; Frechet, Jean M J (2011) Incorporation of carbon nanotubes in porous polymer monolithic capillary columns to enhance the chromatographic separation of small molecules. J Chromatogr A 1218:2546-52
Chambers, Stuart D; Holcombe, Thomas W; Svec, Frantisek et al. (2011) Porous polymer monoliths functionalized through copolymerization of a C60 fullerene-containing methacrylate monomer for highly efficient separations of small molecules. Anal Chem 83:9478-84
Walsh, Zarah; Levkin, Pavel A; Jain, Vijay et al. (2010) Visible light initiated polymerization of styrenic monolithic stationary phases using 470 nm light emitting diode arrays. J Sep Sci 33:61-6

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