Předmět Bioanalytics I - Biomacromolecules (C7942)

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Materiál	Typ	Datum	Počet stažení

Cíl

main objectives of lecture: introduction into bioanalytics; at the end of the course students should be able - to understand approaches in analysis of biomacromolecules and suprabiomolecular structures (nucleic acids, proteins and their complexes, lipidic structures), with special regard to utilisation of mass spectrometry as a versatile analytical tool; - to manage the basic theoretical background for understanding of those approaches

Osnova

analytical approaches in v bioanalytics – demands; high throughput and shotgun analyses, resultaffinity methods – principles, carry-out, use, affinity electrophoresis, IMAC, MOAC, protein equalisersimmunoanalytical methods – principles; immunoanalysis – direct, indirect and immunoassay (FIA, LIA, RIA, EIA, ELISA), IAC, IACE, SELDI…separation of biomacromolecules – gel electrophoresis; principles, carry-out (1D, 2D; polyacrylamide, agarose); blotting; liquid chromatography; principles, carry-out (1D, 2D)basics of mass spectrometry – principles; mass spectrum, basic terminology of MS, ionisation (ESI – spectra deconvolution; MALDI), mass analysis (tandem MS), detection, vacuum techniquegenomics (analysis of nucleic acids) – fundaments, chemical nature of nucleic acids, functional genomics (analytical approaches: PCR, restriction enzymes); identification of known (hybridisation) and unknown sequence (Sanger sequencing, pyrosequencing, 454, Solid); identification of changes in DNA (RFLP, ASO, DGGE, SSCP, CMC); epigenomics; DNA methylation and its analysisproteomics (analysis of proteins and their complexes) – fundamentals, proteome organisation, chemical nature of proteins, their separationexpression proteomics – methodics, protein identification (peptide mapping, MS/MS analysis); quantification (AAA, differential PAGE, MS – direct, indirect, stable isotope labelling); shotgun proteomicsfunctional proteomics – methodics, methods of protein interactions study (qualitative - Y2H; BiFC; mbSUS; SLCA; MeRA; SEAM; ion-mobility strategy; quantitative – MS, QCAT, molar ionisation coefficient; probability approach)structural proteomics – methodics (chemical cross-linking), methods (FTICR MS)post-translational modifications analysis – forms of PTM, MS analysis of PTM – localisation; phosphorylation; PTM extent determinationlipidomics (analysis of lipids and cellular membranes) – fundaments; structure and function of cellular membranes, chemical nature of lipids, analytical approaches (extraction, separation, MS analysis), shotgun lipidomics, lipidomic databasesglycomics (analysis of glycans) – fundaments; chemical nature of glycans and glycoproteins, structure; analytical approaches (derivatisation, separation, MS), shotgun glycomics, glycomic databases

Literatura

Quantitative proteomics by mass spectrometry. Edited by Salvatore Sechi. Totowa, N.J.: Humana Press, 2007. x, 218 p. ISBN 978-1-58829-571-2. infoBENFEY, Philip N. a Alex D. PROTOPAPAS. Essentials of genomics. Upper Saddle River, N.J.: Prentice-Hall, 2005. xiv, 346 p. ISBN 0-13-047018-X. infoMIKKELSEN, Susan R. a Eduardo CORTÓN. Bioanalytical chemistry. Hoboken, N.J.: John Wiley & Sons, 2004. xvii, 361. ISBN 0-471-54447-7. infoLIEBLER, Daniel C. Introduction to proteomics :tools for the new biology. Edited by John R. Yates. Totowa, NJ: Humana Press, 2002. ix, 198 s. ISBN 0-89603-992-7. infoPosttranslational modifications of proteins :tools for functional proteomics. Edited by Christoph Kannicht. Totowa, N.J.: Humana Press, 2002. xi, 322 s. ISBN 0-89603-678-2. info

Požadavky

basic knowledge of biochemistry is prerequisited; mass spectrometry of biomolecules (C7895) is a recommended lecture.

Garant

doc. Mgr. Jan Havliš, Dr.

Vyučující

doc. Mgr. Jan Havliš, Dr.