C7942 Bioanalytics I - Biomacromolecules

Přírodovědecká fakulta
podzim 2019
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.
Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Dodavatelské pracoviště: Národní centrum pro výzkum biomolekul – 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 objective of the course is to introduce the bioanalytics in a form of the group of closely related fields, so-called bio-omics.
Výstupy z učení
at the end of the course students should be able:
to manage basic bioanalytical methods - separation of biomacromolecules including use of affinity, mass spectrometry
to understand principles of basic bio-omics fields (genomics, proteomics, glycomics and lipidomics), including their commonly used bioanalytical approaches and instrumental techniques.
Osnova
  • analytical approaches in v bioanalytics – demands; high throughput and shotgun analyses, result
  • affinity methods – principles, carry-out, use, affinity electrophoresis, IMAC, MOAC, protein equalisers
  • immunoanalytical 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 technique
  • genomics (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 analysis
  • proteomics (analysis of proteins and their complexes) – fundamentals, proteome organisation, chemical nature of proteins, their separation
  • expression proteomics – methodics, protein identification (peptide mapping, MS/MS analysis); quantification (AAA, differential PAGE, MS – direct, indirect, stable isotope labelling); shotgun proteomics
  • functional 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 determination
  • lipidomics (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 databases
  • glycomics (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: 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. Edited by John R. Yates. Totowa, NJ: Humana Press, 2002, ix, 198. ISBN 0896039927. info
  • Posttranslational modifications of proteins : tools for functional proteomics. Edited by Christoph Kannicht. Totowa, N.J.: Humana Press, 2002, xi, 322. ISBN 0896036782. info
Výukové metody
the lecture is based on ppt presentation and its explication. presentation itself 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 of limited availability of textbooks in English covering certain parts of the subject. subject will be eventually lectured if the number of enrolled students exceeds number 5, otherwise students will be given the chance to pass the subject without lecturing combining study materials and on-demand tutorial (at max 2x2h per semester; contact the lecturer to set up the schedule).
Metody hodnocení
oral examination; students are required to understand and be familiar with the principles and its applications. examination consists of three basic questions, which would be during the examination expanded to let the student demonstrate the extent of topic understanding.
Vyučovací jazyk
Angličtina
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Studijní materiály
Předmět je dovoleno ukončit i mimo zkouškové období.
Předmět je vyučován každoročně.
Výuka probíhá každý týden.
Předmět je zařazen také v obdobích podzim 2012, podzim 2013, podzim 2014, podzim 2015, podzim 2016, podzim 2017, podzim 2018, podzim 2020, podzim 2021, podzim 2022, podzim 2023, podzim 2024.