Gram Reaction

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Antigen -Antibody InteractionsPrecipitin Reactions

Hugh B. FackrellHugh B. Fackrell

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Antigen-Antibody Interactions

äAssigned ReadingäAssigned Reading

äContent OutlineäContent Outline

äPerformance OjectivesäPerformance Ojectives

äKey termsäKey terms

äKey ConceptsäKey Concepts

äShort Answer QuestionsäShort Answer Questions

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Assigned Reading

äChapter: 6 pp 144-164äChapter: 6 pp 144-164

äJanis Kuby’s Immunology 3rd EdäJanis Kuby’s Immunology 3rd Ed

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Content Outline

äStrength of Antigen-Antibody InteractionsäStrength of Antigen-Antibody Interactions

äCross ReactivityäCross Reactivity

äPrecipitin ReactionsäPrecipitin Reactions

äAgglutination ReactionsäAgglutination Reactions

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Strength of Antigen-AntibodyInteractions

äaffinityäaffinity

äavidityäavidity

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Cross reactivity

$D:\K06 Ag-Ab reactions\Images\QPT4.gif$

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Mammoth

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Precipitin Reactions

äreactions in fluidsäreactions in fluids

äreaction in gelsäreaction in gels

ä radial immunodiffusionä radial immunodiffusion

ä Ouchterlonyä Ouchterlony

ä immunoelectrophoresisä 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

$D:\K06 Ag-Ab reactions\Images\QPT.gif$

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QPT: Zones

$D:\K06 Ag-Ab reactions\Images\QPT1.gif$

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QPT:# Epitopes

$D:\K06 Ag-Ab reactions\Images\QPT2.gif$

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QPT: Simple system

$D:\K06 Ag-Ab reactions\Images\QPT5.gif$

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QPT: Multiple Interactions

$D:\K06 Ag-Ab reactions\Images\QPT7.gif$

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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 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

ähemagglutinationähemagglutination

äbacterial agglutinationäbacterial agglutination

äpassive agglutinationäpassive agglutination

äagglutination inhibitionäagglutination inhibition

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Hemagglutination

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The End

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Performance Objectives

Key terms, conceptsKey terms, concepts

short answersshort answers

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Key Terms

äagglutination, direct agglutination reaction, indirectagglutination reactionäagglutination, direct agglutination reaction, indirectagglutination reaction

äantibody affinity, antiserum, association constant(K), average affinity,äantibody affinity, antiserum, association constant(K), average affinity,

äaverage intrinsic association constant(Ka), avidity,ELISA, equilibrium constant,äaverage intrinsic association constant(Ka), avidity,ELISA, equilibrium constant,

äequilibrium dialysis, fluorescein, fluorochromes,hemagglutination,äequilibrium dialysis, fluorescein, fluorochromes,hemagglutination,

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äpassive hemagglutination, passivehemagglutination inhibition,äpassive hemagglutination, passivehemagglutination inhibition,

äreverse passive hemagglutination, immuneprecipitation, immunoelectrophoresisäreverse passive hemagglutination, immuneprecipitation, immunoelectrophoresis

äimmunofluorescence, Indirect fluorecentantibody test, ring test,äimmunofluorescence, Indirect fluorecentantibody test, ring test,

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äOuchterlony methods, plasma, primaryantigen-antibody interactions,Radioimmunoassay(RIAäOuchterlony methods, plasma, primaryantigen-antibody interactions,Radioimmunoassay(RIA

äRhodamine, secondary antigen-antibodyinteractions, serology,äRhodamine, secondary antigen-antibodyinteractions, serology,

äserum, titer, zone phenomena (antibodyexcess, antigen excess, equivalence)äserum, titer, zone phenomena (antibodyexcess, antigen excess, equivalence)

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Key Concepts

äExplain a primary antigen-antibody interactionand include at least three importantcharacteristics.äExplain a primary antigen-antibody interactionand include at least three importantcharacteristics.

äDescribe the forces that encourage primaryantigen-Antibody interactionsäDescribe the forces that encourage primaryantigen-Antibody interactions

äAssess the reasons for using the different gelpreciptitin reactionsäAssess the reasons for using the different gelpreciptitin reactions

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äDistinguish betweeen antibody affinity andavidity.äDistinguish betweeen antibody affinity andavidity.

äDescribe the strength of the primaryantigen-antibody interactions usingequilibrium dialysis. Include the terms Kand KaäDescribe the strength of the primaryantigen-antibody interactions usingequilibrium dialysis. Include the terms Kand Ka

äCompare and contrast RIA and ELISAäCompare and contrast RIA and ELISA

äDescribe direct and indirect fluorescentantibody methods.äDescribe direct and indirect fluorescentantibody methods.

äExplain zone phenomena.äExplain zone phenomena.

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äDescribe a secondary antigen-antibodyinteraction in terms of lattice formation andantigen:antibody ratios.äDescribe a secondary antigen-antibodyinteraction in terms of lattice formation andantigen:antibody ratios.

äConstruct a table to compare the variousprocedures used to determine the presence ofsoluble antigen or antibody in a fluid and in agel.äConstruct a table to compare the variousprocedures used to determine the presence ofsoluble antigen or antibody in a fluid and in agel.

äDistinguish between agglutination andpreciptin reactions and give the advantagesand disadvantages of each.äDistinguish between agglutination andpreciptin reactions and give the advantagesand disadvantages of each.

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Short Answer Questions

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äCross reactivity of antibodies createsproblems for their application in serology.Explain.äCross reactivity of antibodies createsproblems for their application in serology.Explain.

äDifferentiate between a primary and asecondary antigen-antibody reaction.äDifferentiate between a primary and asecondary antigen-antibody reaction.

äWhat are three important characteristics thatdistinguish the two reactions?äWhat are three important characteristics thatdistinguish the two reactions?

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äWhat kinds of noncovalent interactions areimportant in antigen-antibody interactions?What aspect of these interactions is mostimportant and why?äWhat kinds of noncovalent interactions areimportant in antigen-antibody interactions?What aspect of these interactions is mostimportant and why?

äHow is equilibrium dialysis used to measurePRIMARY antigen-antibody reactions?äHow is equilibrium dialysis used to measurePRIMARY antigen-antibody reactions?

äDifferentiate between avidity and affinity.äDifferentiate between avidity and affinity.

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äDiscuss the term lattice formation.äDiscuss the term lattice formation.

äWhat are the pros and cons of RIA?äWhat are the pros and cons of RIA?

äDescribe two types of immunofluorescencetests.äDescribe two types of immunofluorescencetests.

äWhat is the advantages of the indirectprocedure over the direct procedure?äWhat is the advantages of the indirectprocedure over the direct procedure?

äWhat are some commonly used fluors?äWhat are some commonly used fluors?

äWhat colour does each fluor emit?äWhat colour does each fluor emit?

äWhat makes precipitin reactions visible?äWhat makes precipitin reactions visible?

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äWhat two factors are important in thedevelopment of precipitin reactions?äWhat two factors are important in thedevelopment of precipitin reactions?

äThree patterns can be observed in theOuchterlony test. DRAW and LABELdiagrams to illustrate these patterns. What doeseach pattern show?äThree patterns can be observed in theOuchterlony test. DRAW and LABELdiagrams to illustrate these patterns. What doeseach pattern show?

äWhat is the major advantage ofimmunoelectrophoresis over immunodiffusion?äWhat is the major advantage ofimmunoelectrophoresis over immunodiffusion?

äWhat are the disadvantages?äWhat are the disadvantages?

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äHow does agglutination differ fromprecipitation?äHow does agglutination differ fromprecipitation?

äWhy are agglutinatin tests more sensitive thatprecipitin tests?äWhy are agglutinatin tests more sensitive thatprecipitin tests?

äDifferentiate between direct and indirectagglutination reactions?äDifferentiate between direct and indirectagglutination reactions?

äWhat is a major advantage of indirectagglutination reaction over direct reactions?äWhat is a major advantage of indirectagglutination reaction over direct reactions?

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Ouchterlony

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Ouchterlony:Partial Identity