26. Monoclonal Antibodies and Their Clinical Applications Flashcards
What is the purpose of the polyethylene glycol (PEG) used to produce hybridomas?
Polyethylene glycol makes membranes fluid so that the cells (B cell and myeloma cell) can fuse together.
How are hybridomas selected for from the mixture of hybridomas, B cells, and myeloma cells during the production of hybridoma cell lines?
Grow cells in a selective media. HAT = only hydridoma cells survive because myeloma cells lack an enzyme needed for growth in this medium but B cells carry it.
How are antigen-specific hybridomas isolated from other hybridomas?
Test culture supernatant from each well for anti-Ag and then grow the positive wells in larger quantities.
How would a hybridoma cell line specific for a human antigen be produced?
- inject human Ag into mouse
- allow time for mouse to develope immune response
- remove B cells from mouse spleen
- fuse with mouse myeloma cells to produce hybridomas and collect monoclonal Ab
How are monoclonal antibodies to lymphocytes used in clinical and research laboratories?
Identification of human lymphocytes via anti-CD markers. - Lymphocytes can be quantitated by immunofluorescence and flow cytometry.
- Typing of leukemias and lymphomas
- Used to monitor transplant patients
Discuss some ways in which monoclonal antibodies are used as therapeutic agents.
Immunotherapy for cancer, autoimmune diseases, allergy, and transplant rejection.
What is the advantage of using a monoclonal antibody as a reagent in a laboratory assay?
- assays are more specific (only against one epitope), but not as sensitive
- homogenous = pure [Decreased lot-to-lot variation)
How are chimeric, humanized, and fully human antibodies different from monoclonal antibodies in terms of their structure? What is the advantage of using these antibodies?
Chimeric = contain mouse variable regoins and human constant regions
Humanized = human Ig containing mouse hypervariable regions
Fully human = derived from human Ig genes introduced into bacteriophages or transgenic mice.
Advantage = lessen possbility of hypersentsitivity eractions to mouse proteins when Abs are used therapeutically.
