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Antigen - Antibody Interactions Precipitin Reactions

Antigen - Antibody Interactions Precipitin Reactions. Hugh B. Fackrell. Antigen-Antibody Interactions. Assigned Reading Content Outline Performance Ojectives Key terms Key Concepts Short Answer Questions. Assigned Reading. Chapter: 6 pp 144-164 Janis Kuby’s Immunology 3rd Ed.

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Antigen - Antibody Interactions Precipitin Reactions

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  1. Antigen -Antibody InteractionsPrecipitin Reactions Hugh B. Fackrell

  2. Antigen-Antibody Interactions • Assigned Reading • Content Outline • Performance Ojectives • Key terms • Key Concepts • Short Answer Questions

  3. Assigned Reading • Chapter: 6 pp 144-164 • Janis Kuby’s Immunology 3rd Ed

  4. Content Outline • Strength of Antigen-Antibody Interactions • Cross Reactivity • Precipitin Reactions • Agglutination Reactions

  5. Strength of Antigen-Antibody Interactions • affinity • avidity

  6. Cross reactivity

  7. Mammoth

  8. Precipitin Reactions • reactions in fluids • reaction in gels • radial immunodiffusion • Ouchterlony • immunoelectrophoresis

  9. Ring Test

  10. Ring Test: Controls

  11. C reactive Protein ring test

  12. QPT

  13. QPT: Zones

  14. QPT:# Epitopes

  15. QPT: Simple system

  16. QPT: Multiple Interactions

  17. QPT: Multiple Reactions

  18. Radial Immuno Diffusion

  19. Double Immuno Diffusion

  20. Immunodiffusion:Single reactions

  21. Immunoelectrophoresis

  22. Counter Immuno Electrophoresis Fig 12-7 Barrett 1988

  23. IEF: Concept

  24. IEF: IgG IgM IgA

  25. IEF: Partial Identity

  26. IEF: Human Serum

  27. Rocket Immunoelectrophoresis

  28. Rocket Immunoelectrophoresis 2

  29. Rocket Immunoelectrophoresis 3

  30. Agglutination Reactions • hemagglutination • bacterial agglutination • passive agglutination • agglutination inhibition

  31. Hemagglutination

  32. The End

  33. Performance Objectives Key terms, concepts short answers

  34. 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,

  35. passive hemagglutination, passive hemagglutination inhibition, • reverse passive hemagglutination, immune precipitation, immunoelectrophoresis • immunofluorescence, Indirect fluorecent antibody test, ring test,

  36. Ouchterlony methods, plasma, primary antigen-antibody interactions, Radioimmunoassay(RIA • Rhodamine, secondary antigen-antibody interactions, serology, • serum, titer, zone phenomena (antibody excess, antigen excess, equivalence)

  37. 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

  38. 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.

  39. 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.

  40. Short Answer Questions

  41. 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?

  42. 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.

  43. 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?

  44. 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?

  45. 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?

  46. Ouchterlony

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