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Lecturer(s)
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Korecká Lucie, doc. RNDr. Ph.D.
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Course content
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Lectures: Immunogens and antigens (immunization, vaccination) Immunoglobulins - structure and function Primary antigen-antibody interaction in solution (non-covalent bond, association constant, affinity and avidity, polyclonal and monoclonal antibodies, genetically engineered antibodies Secondary antigen-antibody interaction in solution and in gels (agglutination reactions, precipitation reactions) Immunoelectrophoretic methods - immunoelectrophoresis according Grabar and Williams, rocket and counter immunoelectrophoresis, two-dimensional (crossed) immuno-electrophoresis, immunofixation Immunoblotting (Western Blots) Immunoassays RIA, EIA Immunofluorescence (FIA, fluorescence-activated-cell-sorting-analysis) Immunoaffinity chromatography (immunomagnetic separation) Isolation and purification of immunoglobulins (non-specific methods, specific methods) practical training in following methods: Serological methods - agglutination reactions, precipitation reactions, immunoturbidimetry Immunodiffusion methods - single and double immunodiffusion Immunoassays - solid-phase - ELISA Electrophoretic separation with protein identification by western blot analysis.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Demonstration, Skills training, Laboratory work
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Learning outcomes
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The aim of this subject is to provide the skills necessary to become an expert in immunochemistry and immunochemical arrays for analysis of food, pathogen identification, toxins and drug estimation. The general goal is to prepare students to the requirements of analytical practice. Laboratory training in a wide range of immunochemical techniques, in their validation and precise interpretation, preparation of reagents and complex kits is also considered part of the educational program.
Student obtain good knowledge of all methods used for analysis of food, methods based on the biospecific reaction between antigen and specific antibody. Student know how to install the test in laboratory, how to optimize the condition and validate the test for commercial application.
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Prerequisites
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Student should know the basics from biology, biochemistry I. and II., microbiology.
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Assessment methods and criteria
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Written examination, Student performance assessment
Written test + oral examination. To take the oral examination, it is necessary to achieve the 60% specified in the written part.
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Recommended literature
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Ferenčík M. Handbook of immunochemistry. Chapman and Hall, London, 1993.
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Johnstone A., Thorpe R. Immunochemistry in practice, Blackwell Science, 1996.
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Phillips T. M. Analytical Techniques in Immunochemistry, Marcel Dekker inc., 1992.
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