1 Antigen Antibody Interactions Precipitin Reactions Hugh B
1 Antigen -Antibody Interactions Precipitin Reactions Hugh B. Fackrell 10/28/202
2 Antigen-Antibody Interactions ä Assigned Reading ä Content Outline ä Performance Ojectives ä Key terms ä Key Concepts ä Short Answer Questions 10/28/202
3 Assigned Reading ä Chapter: 6 pp 144 -164 ä Janis Kuby’s Immunology 3 rd Ed 10/28/202
4 Content Outline ä Strength of Antigen-Antibody Interactions ä Cross Reactivity ä Precipitin Reactions ä Agglutination Reactions 10/28/202
5 Strength of Antigen-Antibody Interactions ä affinity ä avidity 10/28/202
6 Cross reactivity 10/28/202
7 Mammoth 10/28/202
8 Precipitin Reactions ä reactions in fluids ä reaction in gels ä ä ä radial immunodiffusion Ouchterlony immunoelectrophoresis 10/28/202
9 Ring Test 10/28/202
10 Ring Test: Controls 10/28/202
11 C reactive Protein ring test 10/28/202
12 QPT 10/28/202
13 QPT: Zones 10/28/202
14 QPT: # Epitopes 10/28/202
15 10/28/202
16 QPT: Simple system 10/28/202
17 QPT: Multiple Interactions 10/28/202
18 QPT: Multiple Reactions 10/28/202
19 Radial Immuno Diffusion 10/28/202
20 Double Immuno Diffusion 10/28/202
21 Immunodiffusion: Single reactions 10/28/202
22 10/28/202
23 10/28/202
24 Immunoelectrophoresis 10/28/202
25 Counter Immuno Electrophoresis Fig 12 -7 Barrett 1988 10/28/202
26 IEF: Concept 10/28/202
27 IEF: Ig. G Ig. M Ig. A 10/28/202
28 IEF: Partial Identity 10/28/202
29 IEF: Human Serum 10/28/202
30 Rocket Immunoelectrophoresis 10/28/202
31 Rocket Immunoelectrophoresis 2 10/28/202
32 Rocket Immunoelectrophoresis 3 10/28/202
33 Agglutination Reactions ä hemagglutination ä bacterial agglutination ä passive agglutination ä agglutination inhibition 10/28/202
34 10/28/202
35 Hemagglutination 10/28/202
36 The End 10/28/202
37 Performance Objectives Key terms, concepts short answers 10/28/202
38 Key Terms ä agglutination, direct agglutination reaction, indirect agglutination reaction ä antibody affinity, antiserum, association constant (K), average affinity, ä average intrinsic association constant(Ka), avidity, ELISA, equilibrium constant, ä equilibrium dialysis, fluorescein, fluorochromes, hemagglutination, 10/28/202
39 ä passive hemagglutination, passive hemagglutination inhibition, ä reverse passive hemagglutination, immune precipitation, immunoelectrophoresis ä immunofluorescence, Indirect fluorecent antibody test, ring test, 10/28/202
40 ä Ouchterlony methods, plasma, primary antigen-antibody interactions, Radioimmunoassay(RIA ä Rhodamine, secondary antigen-antibody interactions, serology, ä serum, titer, zone phenomena (antibody excess, antigen excess, equivalence) 10/28/202
41 Key Concepts ä Explain a primary antigen-antibody interaction and include at least three important characteristics. ä Describe the forces that encourage primary antigen-Antibody interactions ä Assess the reasons for using the different gel preciptitin reactions 10/28/202
42 ä Distinguish betweeen antibody affinity and avidity. ä Describe the strength of the primary antigen -antibody interactions using equilibrium dialysis. Include the terms K and Ka ä Compare and contrast RIA and ELISA ä Describe direct and indirect fluorescent antibody methods. ä Explain zone phenomena. 10/28/202
43 ä Describe a secondary antigen-antibody interaction in terms of lattice formation and antigen: antibody ratios. ä 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. ä Distinguish between agglutination and preciptin reactions and give the advantages and disadvantages of each. 10/28/202
44 Short Answer Questions 10/28/202
45 ä Cross reactivity of antibodies creates problems for their application in serology. Explain. ä Differentiate between a primary and a secondary antigen-antibody reaction. ä What are three important characteristics that distinguish the two reactions? 10/28/202
46 ä What kinds of noncovalent interactions are important in antigen-antibody interactions? What aspect of these interactions is most important and why? ä How is equilibrium dialysis used to measure PRIMARY antigen-antibody reactions? ä Differentiate between avidity and affinity. 10/28/202
47 ä Discuss the term lattice formation. ä What are the pros and cons of RIA? ä Describe two types of immunofluorescence tests. ä What is the advantages of the indirect procedure over the direct procedure? ä What are some commonly used fluors? ä What colour does each fluor emit? ä What makes precipitin reactions visible? 10/28/202
48 ä What two factors are important in the development of precipitin reactions? ä Three patterns can be observed in the Ouchterlony test. DRAW and LABEL diagrams to illustrate these patterns. What does each pattern show? ä What is the major advantage of immunoelectrophoresis over immunodiffusion? ä What are the disadvantages? 10/28/202
49 ä How does agglutination differ from precipitation? ä Why are agglutinatin tests more sensitive that precipitin tests? ä Differentiate between direct and indirect agglutination reactions? ä What is a major advantage of indirect agglutination reaction over direct reactions? 10/28/202
50 Ouchterlony 10/28/202
51 Ouchterlony: Partial Identity 10/28/202
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