Background
STEP 1 and 2
STEP 3 and 4
STEP 5 and 6
STEP 7
100
In response to an antigen, our immune system produces a heterogeneous mixture of
antibodies
100
What is the animal commonly used?
Mouse
100
Why is the HAT media used?
Because Aminopterin helps block the de novo pathway, and in order for the alternative pathway to take place we need hypoxanthine and thymidine.
100
Common screening techniques
ELISA and RIA
100
What's this step?
Characterization and storing
200
First technique developed for stable monoclonal antibody production
Hybridoma Technology
200
Does the antigen injected to the mouse have different epitopes?
Yes
200
Why are unfused myeloma cells unable to replicate?
Because they have a deficiency in HGPRT
200
Why are hybridomas screened?
For the secretion of the antibody of desired specificity
200
How are hybridomas usually stored?
In liquid nitrogen
300
Derived from single cloned B cells and recognize and bind only to only a particular epitope
Monoclonal antibody
300
Organ to which activated B lymphocytes are harvested
Spleen
300
Why do fused and unfused B cells die shortly?
Because of their short life span
300
What's next after desired hybridomas are identified?
They are isolated and cloned
400
Polyclonal antibodies
Antibodies of different specificities that recognize different epitopes on the same antigen.
They're derived from from multiple clone cells.
400
What are the 5 types of cells produced as a result of cell fusion?
Unfused B-cells, fused B-cells, unfused myelomas, fused myelomas, and hybridomas or hybrid cells.
400
Limiting dilution
Hybridomas are distributed in well culture plates at very low density
400
Do they produce antibodies when they are cloned?
Yes
500
Who described monoclonal antibody production in 1975
Cesar Milstein and Georges Kohler
500
Why are the activated B lymphocytes fused with myeloma cells?
Because activated B lymphocytes can produce antibodies but cannot replicate in culture, and vice versa, myeloma cells cannot produce antibodies but can replicate in culture.
500
Why are hybridomas put in such low density
So that each well can contain one single cell