Agglutination lecture


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  • Ab= anti-SRBC, Ag=SRBC, Step 1= binding of Ab to Ag. Step 2= crosslinking of Ab with 2 Ag to form large aggregates\n
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  • Cheetah:, Bacteria toppings:\n
  • Blue= clinical importance. Since Abs arise in infectious disease and autoimmune diseases. Purple= biological and/or clinical importance. \n
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  • Simple slide is a yes or no as to whether the antiserum contains Abs that are able to agglutinate SRBC (Ag)\n
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  • Excess of Ab- each Ab binds to its own Ag so no cross-linking occurrs\n
  • Anti-SRBC (Rabbit) indicates that the Ab was produced in a rabbit that was injected with sheep red blood cells\n
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  • Agglutination lecture

    1. 1. Agglutination Purpose: To understand thebiological basis for, and applications of, agglutination reactions
    2. 2. Serology: Antibody immunology in vitroSo… the use of antibodies to study theimmune system and stuff.Usually involves Ab-Ag interactions
    3. 3. Pentameric IgM
    4. 4. Agglutination: The action of an antibody when itcross-links multiple antigens producing clumps ofantigens
    5. 5. Agglutination: A Two-Step Process Step Step 1Binding of Ab to Ag 2 Formation of aggregates
    6. 6. Opsinization: the process by which antibody binding to both antigen and phagocyte enhances phagocytosis
    7. 7. *Ab acts as an opsin and makes the bacterium tastierfor the phagocyte AND clumps bacteria togetherallowing the phagocyte to engulf many at once
    8. 8. Importance of measuring Ab-Aginteractions:•Quantifying Ab•Identifying Ab targets•Ab specificity for many targets • Identification • Quantification
    9. 9. Applications of Ab-Ag interactions:•ELISAs•Immunoflorescence•immunodiffusion•Immunoprecipitation•flow cytometry•Western blots•magnetic bead column separation
    10. 10. Simple Slide: Qualitative (visual) confirmation of the specificity of anantiserum to the agglutinated particles. Example: Blood typing
    11. 11. Quantitative determination of the amount of antibody in the serum (titer) No Agglutination Agglutination
    12. 12. Titer: How far you can dilute the patient’s serum and still have it contain enough antibodies to yield a detectable Ab-Ag interaction.*Higher titer means higher starting concentration of Serum/Ab
    13. 13. Using agglutination to track disease progression: What is the titer for this patient?*Dilute the serum, add a constant amount of antigen- You want to know how far you can dilute the patient’s serum (titer) and still have agglutination to determine the concentration of antibody in the original sample
    14. 14. Prozone effect
    15. 15. For today:Sheep Red Blood Cell = SRBC = AgAnti-SRBC antiserum = Ab againstSRBC (rabbit)
    16. 16. Add PBS to wells Add Ab to 1A-12A well 1A Perform serial dilution from well 1A-11A Add Ag to wells 1A-12A*Pipet up and down at least 3 times during serial dilution*Change tip between each serially dilution*Avoid contaminating the Ag stock with Ab!