C8116 Immunochemical techniques

Přírodovědecká fakulta
jaro 2025
Rozsah
2/0/0. 2 kr. (plus ukončení). Ukončení: zk.
Vyučováno kontaktně
Vyučující
Dr. rer. nat. habil. Hans-Heiner Gorris (přednášející)
Garance
Dr. rer. nat. habil. Hans-Heiner Gorris
Ústav biochemie – Chemická sekce – Přírodovědecká fakulta
Dodavatelské pracoviště: Ústav biochemie – Chemická sekce – Přírodovědecká fakulta
Předpoklady
C3181 Biochemie I || C3580 Biochemie || C9530 Strukturní biochemie
A completed bachelor's degree is required for attending the course. Additional recommendation: C3181 Biochemistry I Basic knowledge of biology and general chemistry.
Omezení zápisu do předmětu
Předmět je nabízen i studentům mimo mateřské obory.
Mateřské obory/plány
Cíle předmětu
The course aims at a fundamental understanding of immunochemical methods. The main topics are: A) immune system, B) labelling techniques, C) immunoassays, and D) protein-protein interactions. The course is complementary to C8117 Advanced microscopy and C3002 Nanobiotechnology.
Výstupy z učení
After completing the course, a student will understand - the main players of our immune system and the benefits of getting vaccinated. - what makes antibodies such a terrific biochemical tool. - that immunoassays safe thousands of lifes every day. - how protein-protein interactions hold the key to their function. - what can (and what cannot) be visulized by fluorescence microscopy.
Osnova
  • Introduction: Overview on immunochemical techniques
  • Part A: The immune system
  • 1) General introduction to the immune system
  • 2) Innate / adaptive immune system
  • 3) Lymphoid organs, B cells, clonal selection
  • 4) Generation of antibody diversity, affinity maturation
  • 5) Complement system
  • 6) Immunoglobulin superfamily and function of antibody classes IgG, IgM, IgA, IgE
  • 7) Antibody binding; affinity vs. avidity
  • 8) Antigen determinants: hapten / epitope
  • 9) Raising an immune response in mice, generation of monoclonal antibodies (Milstein/Köhler)
  • 10) role of helper/cytotoxic T cells
  • 11) MHC class I and II, T-cell receptor, antigen presenting cells
  • Part B: Antibodies as immunological tools
  • 1) Handling of antibodies
  • 2) Antibodies as immunochemical reagents
  • 3) Antibody affinity (equilibrium constants, SPR)
  • 5) Limitations of natural antibodies (affinity ceiling, relatively large size)
  • 6) Antibody engineering (excursion: therapeutic antibodies)
  • 8) Alternatives: recombinant, humanized, cameloid, heavy chain antibodies
  • 9) Phage display
  • 10) Aptamers (SELEX)
  • 11) Molecularly imprinted polymers (MIPs)
  • Part C: Immunoassays
  • 1) Definition and key developments of immunoassays
  • 2) Applications of immunoassays (diagnostic, environmental, food safety)
  • 3) Labelling / signal amplification strategies (enzymes, fluorophores, NPs, radionuclides, immune PCR, chemiluminescence)
  • 4) Matrix interference (medical, environmental samples) and non-specific binding
  • 5) Definition: sensitivity / limit of detection / limit of quantification
  • 6) Competitive / non-competitive
  • 7) Heterogeneous / homogeneous immunoassays
  • RIA, ELISA, Time-resolved immunoassays (DELFIA), NP detection (ULISA), fluorescence polarization (FPIA), Fluorescence resonance energy transfer (FRET), proximite ligation
  • 8) Lateral flow assays
  • 9) Biosensors
  • 10) Microarrays (concept of ambient analyte assays)
  • 11) Suspension arrays (magnetic beads)
  • 12) Multiplexing
  • 13) Single-molecule / digital immunoassays (Quanterix Inc.)
  • 14) From our own work: assay design principles
  • 15) Commercialized immunoassay test kits
  • Part D: Immunoaffinity techniques
  • 1) Immune agglutination / precipitation (Nephelometry)
  • 2) Immune diffusion (Ouchterlony test)
  • 3) Immunoblotting
  • 4) Co-immunoprecipitation
  • 5) Analysis of protein-protein interactions
  • (far Western blotting, Affinity chromatography, GST pulldown assay, Tandem affinity purification: tap tagging, analytical ultracentrifugation)
  • 6) Yeast two hybrid system
Výukové metody
Lecture.
Metody hodnocení
Oral examination.
Vyučovací jazyk
Angličtina
Další komentáře
Předmět je dovoleno ukončit i mimo zkouškové období.
Předmět je vyučován každoročně.
Výuka probíhá každý týden.
Předmět je zařazen také v obdobích jaro 2022, jaro 2023, jaro 2024.
  • Statistika zápisu (nejnovější)
  • Permalink: https://is.muni.cz/predmet/sci/jaro2025/C8116