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Lecturer(s)
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Bílková Zuzana, prof. RNDr. Ph.D.
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Course content
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The course follows the knowledge from the field of general biochemistry. It introduces the basics of immunochemistry, describes in a detail the reaction between the antigen and specific antibody and the use of this bioaffinity highly specific and sensitive reaction as a part of analytical or diagnostic tests. Structure of antigen, antibodies, including their preparation (polyclonal, monoclonal, recombinant), production (e.g. vaccination, hybridoma technology, molecular genetic methos), quality control parameters and methods. Students will get acquainted with methods of their isolation and purification (fractionated precipitation, electrophoresis, liquid chromatography, magnetic separation, ultrancentrifugation, etc.) of reagents used for various immunochemical methods. The terms antigenicity, immunogenicity, hapten, determinant, epitope, affinity, avidity will be defined and clarified. Students will familiarize themselves with the principles of IA methods: agglutination reactions, haemagglutination reactions, precipitation, immunodiffusion reactions, immunoelectrophoresis, EIA, RIA, FIA, ECL, blotting techniques, methods of affinity chromatography, immunosensors. Students will be acquainted with principles and arrangement of immunochromatography tests - "clearview test", "rapid test", "bed-side" test, "quick test", etc. Principles of POCT-point-of care tests. Practical examples of the use of IA methods in practice: clinical diagnostics, analysis of food, water, microbiology, environmental analysis, veterinary medicine, etc. Construction of immunoanalytical methods, binding and modification of antibodies, antigens applied in IA tests. New-generation immunonalysts: protein chips, fluidized immunosensors, electrochemical immunosensors, PCD based methods, SELDI etc.
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Learning activities and teaching methods
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Work with text (with textbook, with book), Methods of individual activities
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Learning outcomes
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The subject is based on knowledge from the field of general biochemistry. It introduces the basics of immunochemistry and describes in detail the reaction of an antigen (analyte) with a specific antibody, and the use of this highly specific and sensitive bioaffinity reaction as part of analytical, detection, or diagnostic tests. Structure of antigens, antibodies, including their preparation, production, and quality control. Students will become familiar with the methods of isolation and purification (fractional precipitation, electrophoresis, liquid chromatography, magnetic separation, ultrafiltration, etc.) of reagents used for immunochemical methods. The terms antigenicity, immunogenicity, hapten, determinant, epitope, affinity, and avidity will be explained to students. They will get to know the principles of methods: agglutination reaction, hemagglutination reaction, precipitation, immunodiffusion reaction, immunoelectrophoresis, EIA, RIA, FIA, and ECL, blot technique, affinity chromatography methods, and immunosensors. Students will be introduced to the principles and organization of immunochromatographic tests - clearview, rapid tests, bedside tests, quick tests, etc. Principles of POCT, point-of-care tests. Practical examples of the use of IA methods in practice: clinical diagnostics, analysis of food, water, microbiology, veterinary medicine, etc. Construction of immunoanalytical methods, binding and modification of antibodies and antigens. New-generation immunoanalyzers: protein chips, fluidic immunosensors, electrochemical immunosensors, methods based on SPR, SELDI, etc.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Oral examination, Written examination, Home assignment evaluation
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Recommended literature
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Immunochemical technologies.
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Darwish I.A. .: Immunoassay Methods and their Applications in Pharmaceutical Analysis: Basic Methodology and Recent Advances. Int. J. Biomed. Sci. 2(3) (2006) 217-235.
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Findlay WE a kol. Validation of Immunoassays for Bioanalysis: A Pharmaceutical Industry Perspective. Journal of Pharmaceutical and Biomedical Analysis 2000, 21, 1249-1273.
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Gopinath SCB a kol. Current Aspects in Immunosensors. Biosensors and Bioelectronics 2018, 102, 470-478.
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Hage DS. Handbook of Affinity Chromatography. CRC Press, Taylor and Francis Group, 2005.
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O´Kennedy R, Murphy C. Immunoassays: Developments, Applications and Future Trends. Pan Stanford Publishing: 2017.
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