Antigen - Antibody Interactions Precipitin Reactions

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Antigen -Antibody InteractionsPrecipitin Reactions

Hugh B. FackrellHugh B. Fackrell

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Antigen-Antibody Interactions

Assigned ReadingAssigned Reading Content Outline Content Outline Performance OjectivesPerformance Ojectives

Key termsKey terms Key ConceptsKey Concepts

Short Answer QuestionsShort Answer Questions

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Assigned Reading

Chapter: 6 pp 144-164Chapter: 6 pp 144-164 Janis Kuby’s Immunology 3rd EdJanis Kuby’s Immunology 3rd Ed

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Content Outline

Strength of Antigen-Antibody InteractionsStrength of Antigen-Antibody Interactions Cross ReactivityCross Reactivity Precipitin ReactionsPrecipitin Reactions Agglutination ReactionsAgglutination Reactions

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Strength of Antigen-Antibody Interactions

affinityaffinity avidityavidity

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Cross reactivityCross reactivity

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Mammoth

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Precipitin Reactions

reactions in fluidsreactions in fluids reaction in gelsreaction in gels radial immunodiffusionradial immunodiffusion OuchterlonyOuchterlony immunoelectrophoresisimmunoelectrophoresis

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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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QPTQPT

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QPT: ZonesQPT: Zones

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QPT:# EpitopesQPT:# Epitopes

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QPT: Simple systemQPT: Simple system

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QPT: Multiple InteractionsQPT: Multiple Interactions

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QPT: Multiple ReactionsQPT: 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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ImmunoelectrophoresisImmunoelectrophoresis

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Counter Immuno Electrophoresis

Fig 12-7 Barrett 1988Fig 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

hemagglutinationhemagglutination bacterial agglutinationbacterial agglutination passive agglutinationpassive agglutination agglutination inhibitionagglutination inhibition

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Hemagglutination

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The EndThe End

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Performance ObjectivesPerformance Objectives

Key terms, concepts Key terms, concepts

short answersshort answers

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Key Terms agglutination, direct agglutination reaction, indirect agglutination, direct agglutination reaction, indirect

agglutination reactionagglutination reaction antibody affinity, antiserum, association constant antibody affinity, antiserum, association constant

(K), average affinity, (K), average affinity, average intrinsic association constant(Ka), avidity, average intrinsic association constant(Ka), avidity,

ELISA, equilibrium constant, ELISA, equilibrium constant, equilibrium dialysis, fluorescein, fluorochromes, equilibrium dialysis, fluorescein, fluorochromes,

hemagglutination,hemagglutination,

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passive hemagglutination, passive passive hemagglutination, passive hemagglutination inhibition, hemagglutination inhibition,

reverse passive hemagglutination, immune reverse passive hemagglutination, immune precipitation, immunoelectrophoresisprecipitation, immunoelectrophoresis

immunofluorescence, Indirect fluorecent immunofluorescence, Indirect fluorecent antibody test, ring test,antibody test, ring test,

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Ouchterlony methods, plasma, primary Ouchterlony methods, plasma, primary antigen-antibody interactions, antigen-antibody interactions, Radioimmunoassay(RIARadioimmunoassay(RIA

Rhodamine, secondary antigen-antibody Rhodamine, secondary antigen-antibody interactions, serology, interactions, serology,

serum, titer, zone phenomena (antibody serum, titer, zone phenomena (antibody excess, antigen excess, equivalence)excess, antigen excess, equivalence)

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Key Concepts Explain a primary antigen-antibody interaction Explain a primary antigen-antibody interaction

and include at least three important and include at least three important characteristics.characteristics.

Describe the forces that encourage primary Describe the forces that encourage primary antigen-Antibody interactionsantigen-Antibody interactions

Assess the reasons for using the different gel Assess the reasons for using the different gel preciptitin reactionspreciptitin reactions

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Distinguish betweeen antibody affinity and Distinguish betweeen antibody affinity and avidity.avidity.

Describe the strength of the primary antigen-Describe the strength of the primary antigen-antibody interactions using equilibrium dialysis. antibody interactions using equilibrium dialysis. Include the terms K and KaInclude the terms K and Ka

Compare and contrast RIA and ELISACompare and contrast RIA and ELISA Describe direct and indirect fluorescent antibody Describe direct and indirect fluorescent antibody

methods.methods. Explain zone phenomena.Explain zone phenomena.

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Describe a secondary antigen-antibody Describe a secondary antigen-antibody interaction in terms of lattice formation and interaction in terms of lattice formation and antigen:antibody ratios.antigen:antibody ratios.

Construct a table to compare the various Construct a table to compare the various procedures used to determine the presence of procedures used to determine the presence of soluble antigen or antibody in a fluid and in a soluble antigen or antibody in a fluid and in a gel.gel.

Distinguish between agglutination and Distinguish between agglutination and preciptin reactions and give the advantages preciptin reactions and give the advantages and disadvantages of each.and disadvantages of each.

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Short Answer Questions

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Cross reactivity of antibodies creates Cross reactivity of antibodies creates problems for their application in serology. problems for their application in serology. Explain.Explain.

Differentiate between a primary and a Differentiate between a primary and a secondary antigen-antibody reaction.secondary antigen-antibody reaction.

What are three important characteristics that What are three important characteristics that distinguish the two reactions?distinguish the two reactions?

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What kinds of noncovalent interactions are What kinds of noncovalent interactions are important in antigen-antibody interactions? important in antigen-antibody interactions? What aspect of these interactions is most What aspect of these interactions is most important and why?important and why?

How is equilibrium dialysis used to measure How is equilibrium dialysis used to measure PRIMARY antigen-antibody reactions?PRIMARY antigen-antibody reactions?

Differentiate between avidity and affinity.Differentiate between avidity and affinity.

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Discuss the term lattice formation.Discuss the term lattice formation. What are the pros and cons of RIA?What are the pros and cons of RIA? Describe two types of immunofluorescence tests. Describe two types of immunofluorescence tests. What is the advantages of the indirect procedure What is the advantages of the indirect procedure

over the direct procedure?over the direct procedure? What are some commonly used fluors? What are some commonly used fluors?

What colour does each fluor emit?What colour does each fluor emit? What makes precipitin reactions visible? What makes precipitin reactions visible?

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What two factors are important in the What two factors are important in the development of precipitin reactions?development of precipitin reactions?

Three patterns can be observed in the Three patterns can be observed in the Ouchterlony test. DRAW and LABEL Ouchterlony test. DRAW and LABEL diagrams to illustrate these patterns. What does diagrams to illustrate these patterns. What does each pattern show?each pattern show?

What is the major advantage of What is the major advantage of immunoelectrophoresis over immunodiffusion?immunoelectrophoresis over immunodiffusion?

What are the disadvantages?What are the disadvantages?

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How does agglutination differ from How does agglutination differ from precipitation?precipitation?

Why are agglutinatin tests more sensitive that Why are agglutinatin tests more sensitive that precipitin tests?precipitin tests?

Differentiate between direct and indirect Differentiate between direct and indirect agglutination reactions?agglutination reactions?

What is a major advantage of indirect What is a major advantage of indirect agglutination reaction over direct reactions?agglutination reaction over direct reactions?

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Ouchterlony

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Ouchterlony:Partial Identity