Course will cover nine subjects, each teaching unit will take 3 h. All students must complete additional activities related to regular weekly assignment (2h per student). After each finished unit students in group of three will cover regular weekly assignment connected with the next teaching unit. Microseparation techniques and sample preparation (capillary and nano-techniques applied to sample preparation, purification and concentration, derivatization and sensitive separation). Separation techniques used for quantitative and qualitative analysis and isolation (Capillary electrophoresis, 2-D gel electrophoresis, liquid, ion and immunoaffinity chromatography, Ultracentrifugation and Molecular Characterization and Field-Flow Fractionation). Chemical and enzymatic hydrolysis and proteolysis: Top-down and Bottom-up approach; Methods for protein sequencing (Edman degradation, amino acid analysis, enzymatic cleavages, sequencing in gas-phase, prosthetic groups determination). Posttranslational modifications (Phosphorylation, Glycosylation, Acetylation, Ubiquitination, Disulfide bridges, Sulphatation, Oxidation, Deamidation etc., micro?and macro-heterogeneity and detection methods). Quantitative PCR (qPCR, multiplex qPCR, real-time PCR, reverse transcription quantitative PCR, RT-qPCR). Immunochemical detection methods (Western blot, ELISA, usage of polyclonal and monoclonal antibodies). High-throughput analysis (purpose of HTA, HTA profiling, robotics automation, differential display technique). Bioinformatics and mass spectrometry (peptide and protein databases, search engines and mass spectrometry, MASCOT, ProteinProspector, OMSSA etc)
Students will be able to:
- demonstrate understanding of basic principle of bioanalysis technology assignment, concept, vocabulary and terminology related to macromolecules bioanalysis
- classify biomolecules in relation to separation techniques applications
- list and explain numerous instrumental techniques essential for successful qualitative and quantitative analysis
- prepare suitable samples relative to biomolecule typization and explain biochemical processes and chemical mechanisms related to desired bioanalysis outcome
- resolve complex biopolymer mixture analysis problems in theory and practice
- 1. Microcharacterization of Proteins, Roland Kellner, Friedrich Lottspeich, Helmut E. Meyer, WILLEY VCH, 2nd Edition, 1999.
2. Medicina, Vol.45 No.3, 2009 (http://hrcak.srce.hr/medicina).
3. Principles and practice of bioanalysis, Richard F Venn, 2nd Edition, 2008.