POSITIONS IN ANALYTICAL SYSTEMS BIOLOGY
at the Horváth Laboratory of Bioseparation Sciences, Institute of Analytical Chemistry and Radiochemistry, University of
Innsbruck, Austria
The multiannual support of the European Commission’s Marie Curie Chair and the University of Innsbruck enabled to start a vigorous research
program in the Horváth Laboratory of Bioseparation Sciences to pursue analytical systems biology research, in particular to expand affinity
interaction based bioseparation techniques in genomics, proteomics and glycomics based biomarker discovery using high performance single- and
multi-dimensional bioanalytical tools, novel tagging chemistries and microfabricated device technology connected to mass spectrometry. Current
and prospective research opportunities in analytical systems biology at graduate student and postdoctoral level are as follows:
1. AFFINITY SEPARATION MATERIALS AND BIOMOLECULAR INTERACTION STUDIES
Pioneering the glycoproteomics endeavor in analytical systems biology, our laboratory is developing novel lectin and pseudolectin (boronic acid)
based affinity materials (BLAC) for chromatography and capillary electrophoresis. Multidimensional separation methods, using automated BLAC
technology and microchip liquid chromatography are used to reduce the complexity of biological samples for downstream mass spectrometry analysis.
One of the main goals of this work is to systematically map glycosylation changes of human serum glycoproteins in various cancer types.
2. NOVEL TAGGING CHEMISTRIES FOR QUANTITATIVE PROTEOMICS
Absolute quantification in proteomics is a key issue in gene expression studies at the protein level. To advance quantitative proteomics a new
class of isotope-coded affinity tag reagents are being designed and synthesized in our laboratory, incorporating a fluorescent group making
possible not only relative but absolute quantification of the labeled proteins. FCAT labeling in conjunction with chipLC-MS is used in large
scale quantitative gene expression studies of various human diseases.
3. MICROFABRICATED DEVICE TECHNOLOGY AND CHIP-LC-MS
A high-throughput, miniaturized multicapillary electrophoresis (mCE) approach has been recently introduced by our group for large scale
genotyping and haplotyping. mCE is also being evaluated with a blue light emitting diode (LED) source to profile and sequence complex
carbohydrates released from biologically interesting proteins, in particular to study glycosylation changes associated with cancer. To increase
the resolving power of the process, a miniaturized multidimensional separation approach is being implemented by utilizing BLAC micropipette tips
for glycoprotein fractionation followed fluorophore labeling and mCE analysis.
4. BIOMARKER DISCOVERY AND ANTIBODY PROTEOMICS
Coupling the power of state-of-the-art analytical systems biology with high-throughput monoclonal antibody (mAB) technology, the human plasma
proteome can be adequately queried resulting in discovery and identification of novel protein biomarkers for disease stratification and
monitoring. The antigen of biomarker interest is subject to tryptic digestion using our in-house developed trypsin nanoreactor. The resulting
peptides are then separated by high resolution microchip liquid chromatography and protein identification is accomplished by on-line MS-MS.
This effort focuses on using this global approach to reveal and identify medium and low-abundance plasma protein biomarkers for breast, colon,
lung and prostate cancers.
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