Why is it inefficient to collect antibodies from animals?
They have a polyclonal response to antigen
Summarise the production of monoclonal antibodies
Take B cell producing correct Ab –> fuse with myeloma cell line –> hybridoma –> Ab of interest produced
Describe the production of antibody using recombinant DNA technology
- Isolate genes encoding Ab variable regions
- Construct fusion protein of V region with bacteriophage protein coat
- Clone random populatio of phages to produce phage display library
- Select phage with desired V region and amplify
What is implicated by the suffix of an antibody being “omab”?
mouse monoclonal
What is implicated by the suffix of an antibody being “imab”?
chimeric or partly humanised
What is implicated by the suffix of an antibody being “umab”?
human
Recall 4 therapeutic uses of antibodies
- Cancer drug (herceptin)
- In immunodeficiency
- Blocking cytokine activity
- Removal of T cells from bone marrow grafts
Recall 4 things that can be diagnosed using antibodies
Infection
AI disease
Allergy
Malignancy
Recall 3 antibody diagnostic methods
Tissue typing
Serology
Immunoassays
Describe how ELISA works
2 samples containing Ag used to coat inside of “wells” –> wells covered in Ab against Ag to be measured
If Ag is present, Ab will adhere to well wall -
Reporter is enzyme –> colour change
What is the main use for flow cytometry?
Identify lymphocyte type using Ab against CD markers
What can serum electrophoresis be used to show?
Monoclonal expansion of B cells
Give 3 examples of things ELISA might be used to look for
Tetanus
Hameophillus
Pneumococcus
