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Bioanalytical methods are applied to determination of analytes using specific interactions between molecules. Modern bioanalytical techniques provide high specificity of analysis (one of reagents is biopolymer and that is why we talk about biospecificity), but also high sensitivity and low detection limit. Part of lectures will cover enzymatic methods, proteomics, electromigration methods, genetic methods (particularly PCR), immunochemical and other biochemical and microbiological methods.
Last update: Svoboda Petr (28.05.2020)
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Written examination Last update: Svoboda Petr (28.05.2020)
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R: Králová B., Rauch P., Bioanalytické metody, Vydavatelství VŠCHT Praha, 2. vydání 1995, ISBN 80-7080-449-1 R: Králová B., Rauch P., Fukal L., Bioanalytická chemie, Vydavatelství VŠCHT Praha, 2007, ISBN 80-7080-449-1 R: Káš J., Kodíček M., Valentová O., Laboratorní techniky biochemie, Praha 2005, ISBN 80-7080-586-2 R: Ruml T., Rumlová M., Pačes V., Genové inženýrství, Praha 2002, ISBN 80-7080-499-8 R: Daussant J., Desvaux F.-X., Introduction to immunochemical techniques for medical diagnosis, food quality control and environmental testing, 2007, ICT Press, 9788070806418 Last update: Kubová Petra (18.12.2017)
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1. Definition of bioanalytical methods as methods based on biological recognition; Basics of molecules isolation from a biological material; Stability of biological materials.
2. Enzymes 1: Enzymes as analytical agents - introduction, kinetics of enzyme reactions; Methods for determination of substrates, activators and inhibitors of enzyme reactions.
3. Enzymes 2: Enzymes used in free and immobilized forms; Application of enzymatic methods in clinical and experimental practice.
4. Immunochemical methods 1: Principles of immunochemical methods - antigen, hapten, antibodies; Characterization of antigen-antibody interactions; Principles of the immune response; Antibodies - preparation, usage, polyclonal, monoclonal, recombinant.
5. Immunochemical methods 2: Classification of immunochemical methods; Immunoprecipitation methods and their use in practice - precipitation in solution, agglutination and passive agglutination; Sensitive immunochemical techniques (ELISA); Bioanalytical methods using radionuclides (suitable radionuclides, half-lives, detection methods, antibody labelling), RIA, radiometric methods, work with radionuclides.
6. Electrophoretic methods 1: Principles of electrophoretic methods - SDS electrophoresis, isoelectric focusing, capillary electrophoresis; immunoelectrophoretic methods.
7. Electrophoretic methods 2: Blotting techniques - western blot - detection, isolation and characterization of proteins after the electrophoretic separation, staining and chemiluminescence detection; Northern blot; Southern blot.
8. Genetic methods 1: Polymerase chain reaction (PCR), digital PCR, quantitative PCR, in situ detection of specific product sequences.
9: Genetic methods 2: DNA and RNA array; DNA sequencing; Application of genetic methods.
10. In vitro techniques: Use of cell lines in bioanalytical methods; Biochemical and microbiological methods in analytical chemistry.
11. Chromatographic techniques 1: LC, GC, bioaffinity chromatography, affinity precipitation, immunoaffinity biosensors.
12. Chromatographic techniques 2: Application of chromatographic techniques; Flow cytometry.
13. Mass spectrometry of biomolecules.
14. Detection limits and application of selected methods; Usage of modern analytical methods for biopolymers. Last update: Svoboda Petr (28.05.2020)
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Additional list of suggested bibliography and electronic references will be provided within individual lectures. Last update: Kubová Petra (18.12.2017)
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Students will be able to: Choose suitable analytical method with appropriate detection limit. Apply the chosen method for samples of different origin. Orientate themselves in specialized literature concerning biomolecules issue. Obtain wide knowledge of bioanalytical methods spectra including possibility and limits of their application. Last update: Kubová Petra (18.12.2017)
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Biochemistry I Last update: Svoboda Petr (18.11.2021)
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Teaching methods | ||||
Activity | Credits | Hours | ||
Účast na přednáškách | 1 | 28 | ||
Příprava na zkoušku a její absolvování | 2 | 56 | ||
3 / 3 | 84 / 84 |