Bi7940 Bioanalytics I - Biomacromolecules

Přírodovědecká fakulta
podzim 2008
Rozsah
2/0/0. 2 kr. (plus ukončení). Doporučované ukončení: zk. Jiná možná ukončení: k.
Vyučující
doc. Mgr. Jan Havliš, Dr. (přednášející)
Garance
doc. Mgr. Jan Havliš, Dr.
Ústav experimentální biologie – Biologická sekce – Přírodovědecká fakulta
Předpoklady
basic knowledge of biochemistry is prerequisited; mass spectrometry of biomolecules (C7895) is a recommended lecture.
Omezení zápisu do předmětu
Předmět je nabízen i studentům mimo mateřské obory.
Mateřské obory/plány
Cíle předmětu
main objectives of lecture: introduction into bioanalytics; understanding the approaches in analysis of biomacromolecules snd suprabiomolecular structures (nucleic acids, proteins and their complexes, lipidic structures), with special regard to utilisation of mass spectrometry as a versatile analytical tool; managing the important theoretical background for understanding of those approaches
Osnova
  • 1) analytical approaches in v bioanalytics – demands; high throughput, shotgun analysis, standardization
  • 2) affinity methods – principles, carry-out and use (separation, interaction studies), IMAC, APAGE
  • 3) immunoanalytical methods – principles; immunoanalysis – precipitation, agglutination, immunoassay, methods: FIA, LIA, RIA, EIA, ELISA, IAC, IACE, SELDI…
  • 4) separation of biomacromolecules – gel electrophoresis; principles, carry-out and use (1D, 2D; polyacrylamide, agarose); blotting; 2D liquid chromatography
  • 5) genomics (analysis of DNA) – fundaments of genetics (structure, function of DNA), analytical approaches: PCR, restriction enzymes; identification of known (hybridisation) and unknown sequence (Sanger m., pyrosequencing, 454, Solid); identification of changes in DNA; DNA methylation and its analysis
  • 6) basics of mass spectrometry – principles; ionisation (ESI –spectra deconvolution; MALDI), mass analysis (tandem MS), detection, vacuum technique mass spectrum, basic terminology of MS
  • 7) proteomics (analysis of proteins and their complexes) – fundamentals (structure of proteins and protein complexes, proteome organisation)
  • 8) expression proteomics – methodics (MS: bottom-up, top-down), MS: identification of protein using peptide fingerprints/maps, bioinformatics in MS-assisted proteomics; MS: identification of protein using amino acid sequence; other methods of expression proteomics: 2D HPLC, 2D PAGE, 2D imaging
  • 9) quantitative analysis of protein complexes – quantification using mass spectrometry, input of quantitative information into MS spectrum (internal standard method; mass tagging – ICAT, ICMT, TMT, MCAT, GIST, SILAC, SIL), deciphering the quantitative information in MS spectrum; automation in proteomics (AI problem)
  • 10) functional proteomics – methodics (parallel and sequential analysis), methods of protein-protein interaction study (Y2H; BiFC; mbSUS; MeRA; SEAM –myc, TAP, FLAG, His-tags; ion-mobility strategy); absolute quantification of proteins and peptides by MS (SIL, VICAT, QCAT; molar ionisation coefficient; probability approach)
  • 11) structural proteomics – methodics (chemical cross-linking), methods (FTICR MS); shotgun proteomics – hyphenated techniques 12) post-translational modifications analysis – forms of PTM, MS analysis of PTM – localisation; glycosylation, phosphorylation; PTM extent determination
  • 13) lipidomics (analysis of lipids and cellular membranes) – fundamentals (structure and function of cellular membranes and their components), analytical approaches: lipid extraction, TLC, GC, LC, SFC, GPC, MS analysis (PIS, HGS, FAS), shot-gun analysis
Literatura
  • MUSHEGIAN, Arcady R. Foundations of comparative genomics. 1st ed. Burlington: Elsevier Academic Press, 2007, 265 s. ISBN 9780120887941. info
  • Quantitative proteomics by mass spectrometry. Edited by Salvatore Sechi. Totowa: Humana Press, 2007, x, 218. ISBN 9781588295712. info
  • BENFEY, Philip N. a Alex D. PROTOPAPAS. Essentials of genomics. Upper Saddle River: Prentice-Hall, 2005, xiv, 346. ISBN 013047018X. info
  • MIKKELSEN, Susan R. a Eduardo CORTÓN. Bioanalytical chemistry. Hoboken, N.J.: John Wiley & Sons, 2004, xvii, 361. ISBN 0471544477. info
  • LIEBLER, Daniel C. Introduction to proteomics : tools for the new biology. Totowa, NJ: Humana Press, 2002, ix, 198 s. ISBN 0-89603-991-9. info
  • Posttranslational modifications of proteins : tools for functional proteomics. Edited by Christoph Kannicht. Totowa, N.J.: Humana Press, 2002, xi, 322. ISBN 0896036782. info
Metody hodnocení
the lecture is based on ppt presentation and its explication, presentation its-self will be available as a study material (black-and-white printable pdf with high resolution and restricted access rights). it is recommended to attend the lecture, because of the explication, which significantly extends the presentation and because there are no available textbooks covering the subject.
oral examination; students are required to understand and be familiar with the principles and its applications; enumerations, listing of facts and suchs would not be required.
Vyučovací jazyk
Angličtina
Další komentáře
Předmět je dovoleno ukončit i mimo zkouškové období.
Předmět je vyučován každoročně.
Výuka probíhá každý týden.
Předmět je zařazen také v obdobích podzim 2010 - akreditace, podzim 2009, podzim 2010, podzim 2011, podzim 2011 - akreditace.