Gram Reaction
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Transcript Gram Reaction
1
Antigen -Antibody Interactions
Precipitin Reactions
Hugh B. Fackrell
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Antigen-Antibody Interactions
Assigned Reading
Content Outline
Performance Ojectives
Key
terms
Key Concepts
Short Answer Questions
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Assigned Reading
Chapter: 6 pp 144-164
Janis Kuby’s Immunology 3rd Ed
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Content Outline
Strength of Antigen-Antibody Interactions
Cross Reactivity
Precipitin Reactions
Agglutination Reactions
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Strength of Antigen-Antibody
Interactions
affinity
avidity
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Cross reactivity
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Mammoth
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Precipitin Reactions
reactions in fluids
reaction in gels
radial immunodiffusion
Ouchterlony
immunoelectrophoresis
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Ring Test
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Ring Test: Controls
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C reactive Protein ring test
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QPT
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QPT: Zones
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QPT:# Epitopes
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QPT: Simple system
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QPT: Multiple Interactions
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QPT: Multiple Reactions
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Radial Immuno Diffusion
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Double Immuno Diffusion
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Immunodiffusion:
Single reactions
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Immunoelectrophoresis
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Counter Immuno Electrophoresis
Fig 12-7 Barrett 1988
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IEF: Concept
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IEF: IgG IgM IgA
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IEF: Partial Identity
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IEF: Human Serum
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Rocket Immunoelectrophoresis
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Rocket Immunoelectrophoresis 2
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Rocket Immunoelectrophoresis 3
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Agglutination Reactions
hemagglutination
bacterial agglutination
passive agglutination
agglutination inhibition
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Hemagglutination
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The End
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Performance Objectives
Key terms, concepts
short answers
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Key Terms
agglutination, direct agglutination reaction, indirect
agglutination reaction
antibody affinity, antiserum, association constant
(K), average affinity,
average intrinsic association constant(Ka), avidity,
ELISA, equilibrium constant,
equilibrium dialysis, fluorescein, fluorochromes,
hemagglutination,
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passive hemagglutination, passive
hemagglutination inhibition,
reverse passive hemagglutination, immune
precipitation, immunoelectrophoresis
immunofluorescence, Indirect fluorecent
antibody test, ring test,
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Ouchterlony methods, plasma, primary
antigen-antibody interactions,
Radioimmunoassay(RIA
Rhodamine, secondary antigen-antibody
interactions, serology,
serum, titer, zone phenomena (antibody
excess, antigen excess, equivalence)
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Key Concepts
Explain a primary antigen-antibody interaction
and include at least three important
characteristics.
Describe the forces that encourage primary
antigen-Antibody interactions
Assess the reasons for using the different gel
preciptitin reactions
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Distinguish betweeen antibody affinity and
avidity.
Describe the strength of the primary
antigen-antibody interactions using
equilibrium dialysis. Include the terms K
and Ka
Compare and contrast RIA and ELISA
Describe direct and indirect fluorescent
antibody methods.
Explain zone phenomena.
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Describe a secondary antigen-antibody
interaction in terms of lattice formation and
antigen:antibody ratios.
Construct a table to compare the various
procedures used to determine the presence of
soluble antigen or antibody in a fluid and in a
gel.
Distinguish between agglutination and
preciptin reactions and give the advantages
and disadvantages of each.
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Short Answer Questions
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Cross reactivity of antibodies creates
problems for their application in serology.
Explain.
Differentiate between a primary and a
secondary antigen-antibody reaction.
What are three important characteristics that
distinguish the two reactions?
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What kinds of noncovalent interactions are
important in antigen-antibody interactions?
What aspect of these interactions is most
important and why?
How is equilibrium dialysis used to measure
PRIMARY antigen-antibody reactions?
Differentiate between avidity and affinity.
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Discuss the term lattice formation.
What are the pros and cons of RIA?
Describe two types of immunofluorescence
tests.
What is the advantages of the indirect
procedure over the direct procedure?
What are some commonly used fluors?
What
colour does each fluor emit?
What makes precipitin reactions visible?
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What two factors are important in the
development of precipitin reactions?
Three patterns can be observed in the
Ouchterlony test. DRAW and LABEL
diagrams to illustrate these patterns. What does
each pattern show?
What is the major advantage of
immunoelectrophoresis over immunodiffusion?
What are the disadvantages?
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How does agglutination differ from
precipitation?
Why are agglutinatin tests more sensitive that
precipitin tests?
Differentiate between direct and indirect
agglutination reactions?
What is a major advantage of indirect
agglutination reaction over direct reactions?
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Ouchterlony
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Ouchterlony:Partial Identity
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