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Immunology Chapter 6, Lecture 1. Richard L. Myers, Ph.D. Department of Biology Southwest Missouri State Temple Hall 227 Telephone: 417-836-5307 Email: rlm967f@mail.smsu.edu Homepage: http://creative.smsu.edu/biology/myersr/index.html TopClass: http://creative.smsu.edu.

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immunology chapter 6 lecture 1
ImmunologyChapter 6, Lecture 1
  • Richard L. Myers, Ph.D.
  • Department of Biology
  • Southwest Missouri State
  • Temple Hall 227
  • Telephone: 417-836-5307
  • Email: rlm967f@mail.smsu.edu
  • Homepage: http://creative.smsu.edu/biology/myersr/index.html
  • TopClass: http://creative.smsu.edu
antigen antibody interactions
Antigen-Antibody interactions
  • Similar to enzyme-substrate reactions
  • Involves noncovalent interactions
    • between epitope and CDRs of antibody
    • the reaction is very specific
      • used in diagnosing disease
      • monitoring level of the humoral response
      • identifying certain molecules
  • Ag-Ab reactions differ and speed and sensitivity
strength of ag ab reactions
Strength of Ag-Ab reactions
  • Involves noncovalent interactions
    • hydrogen bonds
    • ionic bonds
    • hydrophobic interactions
    • van der Waals interactions
  • Requires many of these interactions
    • operating over a small distance (1 angstrom)
  • Therefore, depends upon a very close fit
antibody affinity
Antibody affinity
  • Defined as the strength of the sum total of noncovalent interactions
  • Low-affinity antibodies bind antigen weakly
  • High-affinity bind more tightly
  • Ag-Ab association can be determined by equilibrium dialysis
antibody avidity
Antibody avidity
  • Defined as the strength of multiple interactions between multivalent antibody and antigen
  • IgM has a low affinity compared to IgG, but has a high avidity
cross reactivity
Cross-reactivity
  • Ag-Ab reactions are highly specific, but
    • Ab may cross-react with an unrelated Ag
    • affinity is usually less
  • An example of a cross-reacting Ab is the ABO blood-group antigens
    • originally elicited by microbial antigens
    • will cross-react with similar antigens on RBCs
precipitin reactions
Precipitin reactions
  • Results when antigen and antibody interact
  • Forms a lattice that becomes visible
    • antibody must be bivalent
    • called precipitins
  • Reaction may take place in fluids
    • dilute the antibody, hold antigen constant
    • used as a rapid test for presence of Ag or Ab
      • the interfacial or ring test is very useful
precipitin reactions cont
Precipitin reactions (cont)
  • Reactions may occur in gels
    • produces a visible line of precipitation
    • called immunodiffusion
      • used to determine relative concentration of Ag or Ab
      • used to compare antigens
      • or determine relative purity of an antigen
  • Mancini method (radial diffusion)
    • antigen placed in a well
    • diluted antibody placed into agar
precipitin reactions cont1
Precipitin reactions (cont)
  • Ouchterlony method
    • double diffusion in agar
    • antigen and antibody diffuse toward each other
    • form visible precipitation line where they meet
    • show relationships between different systems
      • identity
      • nonidentity
      • partial identity
precipitin reactions cont2
Precipitin reactions (cont)
  • Immunoelectrophoresis
    • seperation of proteins with electrophoresis
    • followed by double diffusion in gel
    • can detect concentrations of 3 - 20 mg/ml
    • rocket electrophoresis is similar but can detect level of protein as low as 0.2 mg/ml
  • 2-D immunoelectrophoresis
    • electrophoresis followed by overlay with gel with antiserum
assignment
Assignment
  • Read Chapter 6, Antigen-Antibody Interactions
  • Review question 6 (pg 163)