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3 D Structure of Immunoglobulins

3 D Structure of Immunoglobulins. Hugh B. Fackrell Filename:Kuby05b7.ppt. Assigned Reading Content Outline Performance Objectives Key terms Key Concepts Short Answer Questions. ASSIGNED READING. Chapter pp. Antibody structure. Special antibodies to study structure Myeloma proteins

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3 D Structure of Immunoglobulins

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  1. 3 D Structure of Immunoglobulins Hugh B. Fackrell Filename:Kuby05b7.ppt

  2. Assigned Reading • Content Outline • Performance Objectives • Key terms • Key Concepts • Short Answer Questions

  3. ASSIGNED READING • Chapter pp

  4. Antibody structure • Special antibodies to study structure • Myeloma proteins • Bence Jones Proteins • Monoclonal Antibodies • 3D structure of antibodies • Binding site • Variability in structure

  5. Angels & Devils

  6. EM of IgA

  7. Antibody flexibility

  8. Antibody: Space filling model

  9. Mulptiple Myeloma • Disease • Plasmocytoma • Cancer of antibody producing cell • Symptoms • recurrent infections, anaemia, fractures, pain, bone lesions • Detection • Electrophoresis, X-rays, blood smear

  10. IgG Crystal

  11. Myeloma Induction

  12. Bence Jones Protein • Homogenous light chains • In urine of myeloma patients • Easily purified • Insoluble at 60 -70 oC • Soluble at 80 oC

  13. Monoclonal Antibodies • Add information here

  14. Antibody model

  15. Problems with flexibility • Difficult to determine entire structure • Hinge flexibility • Fragment • Study Fragment • Assemble composite

  16. Hinge Prolines

  17. IgG: Enzyme hydrolysis

  18. Bence Jones Light chains • Simplest fragments to isolate • Easily crystallized • Mimics Fab

  19. Bence Jones Monomer

  20. Bence Jones Dimer

  21. Immunoglobulin fold • Signature structure common to all immunoglobulins • 2 beta sheets • held by disulfide bond • Variations in detailed structure set specificity

  22. Ig Fold: Variable domains

  23. Ig fold: Constant domains

  24. Antigen Binding Site

  25. Structure of IgG

  26. Ig Fold: hypervariable regions

  27. Vitamin K bound to IgG

  28. Antigen binding to Antibody

  29. Antibody Strucutre: Internal motion

  30. Antibody Structure: Internal movement

  31. Antigen in Antibody

  32. Hypervariable Regions • Discovery • KW plots • Light chains • Heavy chains • CDR vs HVR

  33. Kabat & Wu Plots • Why • demonstrate details of variability of protein sequence of similar proteins • Results • Variability = 1.0 >> no variation • Variability = 400 >> 20 different AA Number different amino acids Variability = Frequency of most common AA

  34. HV: light chains

  35. KW plot heavy chains

  36. HV:IgG

  37. KW Heavy chains CDR

  38. KW Light chains CDR

  39. Ig Superfamily

  40. Isotypes Allotypes Idiotypes

  41. Allosera Production

  42. Isotypes, Allotype Idiotype

  43. The End

  44. Performance Objectives Key terms, concepts short answers

  45. SHORT ANSWER QUESTIONS

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