Mgr. Jana Křenková, Ph.D.

Project: NanoBioSep - Nanoparticle modified monolithic supports for bioanalysis

Person in Charge: Ing. František Foret, CSc.

Host institution: Institute of Analytical Chemistry of the AS CR

Country of Origin: Czech Republic

Country of scientific activity: USA

Project duration: 24 months

Scientific panel: Chemistry

Abstract:

The objective of this project is to develop methodology for synthesis of metal oxide nanoparticles and their immobilization on the pore surface of organic polymer monolith in order to obtain new types of chromatographic media affording improved retention of isolated/separated compounds, lower non-specific interaction, and higher stability. Specifically, the effort will be focused on the metal oxide nanoparticles such as iron oxide and nickel oxide nanoparticles synthesized using various wet chemical methods such as co-precipitation and thermal decomposition. The methacrylate based monolithic supports will be prepared by thermally and UV initiated polymerization in fused silica capillaries and the metal oxide nanoparticles will be immobilized on their pore surfaces using various chemistries. The effect of particle size as well as morphology and chemistry of monolithic support on the nanoparticle immobilization and chromatographic performance of developed columns will be studied in detail. The complexity of biological materials, especially when dealing with protein analysis, always requires the use of multidimensional analysis for identification and quantification of individual components. Therefore, these new types of monolithic capillary columns will be integrated into miniaturized system combining sample pretreatment, separation and mass spectrometry detection, and applied for analysis of proteins and selective enrichment of phoshopeptides. The main advantage of such a system, compared to off-line arrangement, will be speed of analysis, small sample consumption, and minimization of sample loss and contamination.

The ongoing project summary to be published

• During the first year of the project, I have been working on the synthesis of iron oxide nanoparticles and their incorporation into the organic polymer-based monolithic materials prepared in various formats (capillary column, pipette tips). The developed materials were characterized by several techniques such as dynamic light scattering or scanning electron microscopy and successfully applied for phosphoproteomic analysis.
I have also been involved in the project, which is focused on the development of new instrumentation for on-line coupling of capillary electrophoresis with mass spectrometry. The hybrid capillary/microfluidics device was designed, fabricated and successfully applied for on-line CE/MS analyses of peptides, proteins, and oligosaccharides.
In collaboration with the Department of Liquid-Phase Separations of the Institute of Analytical Chemistry of the ASCR, v. v. i., a divergent flow isoelectric focusing device has been applied for the separation of peptides. The potential of the developed device for preparative scale separations was demonstrated by isolation of highly purified caseinomacropeptides used as ingredients in diets designed for people suffering from phenylketonuria.

• The obtained results have been presented at the international conferences (HPLC2011, Budapest, Hungary; CECE2011, Brno, Czech Republic and HPLC2012, Anaheim, USA) and published in the scientific journals with high impact factors (Electrophoresis, Journal of Separation Science, Proteomics, Analytical and Bioanalytical Chemistry). The poster entitled “Nanoparticle-modified monolithic pipette tips for selective enrichment of phosphopeptides” was awarded as one of the ten best posters of the HPLC2012 conference where more than 400 posters were presented.

• In 2011, my research work was honored with the Otto Wichterle Award, an honor for young researches given by the Academy of Sciences of the Czech Republic.

• I have also established several local and international collaborations with the research groups from the Czech Republic, Hungary or USA, which led to joint publications or presentations at the scientific conferences.