Antibody ID Special Techniques

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Antibody ID Special Techniques
2013-04-21 16:00:15
Antibody ID Techniques

Antibody ID Special Techniques
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  1. What is Titration
    A method to determine the highest dilution at which agglutination can be detected.
  2. When is titration used?
    • Prenatal Studies:
    •         - Monitors maternal IgG antibody
    • Antibody ID for: 
    •         - CAS
    •         - HTLA
  3. What is a signifcant titration result for HDFN?
    Fourfold rise or greater
  4. Titration with CAS are what?
    • Pathologic and have high titers (>64).
    • React at 37C.
  5. HTLA with titration techniques are what?
    • Weakly reactive IgG antibodies that are not clinically significant byt may mask other antibodies.
    • React with high dilutions
  6. What are the antibodies seen with HTLA?
    • Ch
    • Rg
    • Kna  
    • McCa
    • Csa

  7. Titration procedure is?
    • Patient serum diluted with diluent (saline, albumin, etc.)
    • Incubate at 37C & AHG for IgG
    • Incubate at room temp & 4C for IgM
    • Last dilution with a reaction is result.
  8. Neutralization is?
    Inactivation of an antibody resulting from binding with a soluble antigen.
  9. Known substances that have blood group antigen specificity for:
    Hydatid cyst fluid-
    P1 Substance
  10. Known substances that have blood group antigen specificity for:
    Human Urine
  11. Known substances that have blood group antigen specificity for:
    Human Serum/plasma
    • Ch
    • Rg
    • Lewis System
  12. Known substances that have blood group antigen specificity for:
    Human Breast milk
  13. Neutralization aplications for antibody ID is & procedure
    Multiple antibodies - Neutralize one antibody and allow for the detection of another underlying antibody.

    • Mix patient serum with antigen containing substance.
    • If present antibody binds to substance
    • Treated serum is then tested with known antigen pos cells
  14. Adsorption Definition:
    • Remove antibody from a serum sample by combining it with antigen positive red cells.
    • ie.
    • Serum sample with anti-A is incubated with group A red cells. Then anti-A will attach to (adsorb onto) the  red cells leaving the serum non-reactive for the anti-A.
  15. Adsoption studies are useful for?
    • -Separate multiple antibodies
    • -Remove autoantibody in order to identify alloantibody.
    • Frequently used fro WAIHA once elution doesn't work.
    • Confirm the specificity of an antibody by showing it can be adsorbed onto a red cell of a known antigen type.
  16. How to rule out Alloantibodies from adsorption?
    Must adsorb the autoantibody from the serum and then test the adsorbed serum for alloantibodies.
  17. What is Autologous adsoption?
    • Is used when patient has NOT been recently transfused.
    •        - Patient own cells are treated with ZZAP to prepare cells. Trying to destroy autoantibody. ONLY auto will be on patient cells.
    • Now treated cells are incubated with the patients serum.
  18. What is Homologous/allogeneic Adsorption?
    • Used for patients who have recently been transfused. So, you can't use auto cells becuase allo may be adsorbed.
    • Use red cells of known phenotype to adsorb out the broad specificity auto.
    • Use typically 3 different cells:
    • R1R1, R2R2, rr lacking Jka, Jkb, K & s antigens. 
    • Enzyme Treat to make them Fya, Fyb, & S antigen neg.
  19. What is Elution?
    • Is a technique used to free the antibody bound to the red cell membrane. By either:
    • Temperature for: 
    •     - ABO system antibodies
    • Sovents for:
    •     - Investigation of warm reactive allo or autoantibodies.
    • Investiage a pos DAT
  20. When is it useful to use both Adsorption and Elution techniques?
    Separate multiple antibodies (frequently seen with WAIHA) and then test to find allo.