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Lecturer(s)
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Bílková Zuzana, prof. RNDr. Ph.D.
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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 Purification and analysis of IgG - non-specific methods (fractionation precipitation, electrophoretic methods, DEAE anion-exchange chromatography, gel filtration chromatography, SDS-PAGE analysis of IgG subunit structure - MR,) Preparation of pure immunoglobulin - affinity chromatography of antibodies
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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 develop 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 develop the student in a flexible fashion adapted 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 cellular biology, microbiology and immunology. C536 Food biochemistry
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Assessment methods and criteria
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Written examination, Student performance assessment
written test
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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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