Immunologic Tests Flashcards
The process of forming monoclonal Ab involves fusing what cells?
Mouse antibody producing B cells Human hybridomas (myeloma)
When growing hybridomas on a HAT plate, what is the goal?
To select for cells that are truly B hybrids with myeloma cells
What is the HAT plate made of?
Hypoxanthine, aminopetran and thymidine
What does hypoxanthine and thymidine’s role in a HAT plate?
Myeloma cannot survive with only hypoxanthine and thymidine. B cells cannot survive for two weeks, but CAN survive with hypoxanthine and thymidine. When fused together, they can both survive.
What does aminopterin do?
Blocks synthesis of TTP and GTP through dihydrofolate reductase
What is the difference between direct and passive agglutination assays?
Direct: Directly seeing interaction between antibody and cell
Passive: Antibody linked to something you can see (Latex beads)
If a cell lives in a neutralization assay, did neutralization occur?
Yes
What is the difference between direct and indirect immunofluorescence?
Direct: Only one antibody attached to both active site and GFP
Indirect: One antibody attached to active site, one antibody attached to that antibody and GFP
In a direct ELISA test, what do you start out with on the plate?
Antibodies
In a direct ELISA test, what is the order of adding?
- Patient sample
- Antivirus antibody containing conjugated enzyme
- Wash
- Add substrate for enzyme, and measure colored product
In an indirect ELISA test, what do you start off with on the plate?
Antigen
In an indirect ELISA test, what is the order of adding?
- Serum
- Wash
- anti-IgG antibodies conjugated to enzyme
- Wash
- Substrate for enzyme, measure colored product
In a competitive ELISA test, what do you start out with in the well?
Antigen with known amount of antigen specific antibody
In a competitive ELISA test, what is the order of adding?
- Antibody/specimen complex
- Wash
- Anti-IgG antibodies conjugated to enzymes
- Wash
- Add substrate, measure amount of colored
What is different about reading a competitive ELISA as compared to direct/indirect?
The more color, the less antigen.
