L5_Immunoassays Flashcards
Define Avidity
The strength of binding of multivalent antiserum to multivalent antigen
what are the two main techniques used to eliminate cross-reactivity
absorption and affinity chromatography
Describe Monoclonal Antibody Affinity, Avidity and cross reactivity compared to polyclonal
They have lower cross reactivity, lower affinity and little avidity.
What is a myeloma cell?
A tumor of a plasma cell that no longer creates antibodies by has the capability to do so.
Describe the process of making a monoclonal antibody
- Immunize an animal
- Isolate spleen cells form immunized animal
- Fuse spleen cells to plasmacytoma tumor cells (myeloma)
- Select for only those cells that are hybrids
- Clone hybridomas so each single cell grows up independently
- selelct the individual clone with the specificity you are interested in
What will a drug name end with if it is a monoclonal antibody
mab
What is the function of framework residues?
These residues are not directly involved in antigen binding but instead are essential in producing the folding of the V-region producing the antigen binding site.
Describe the technique of absorption
Absorption uses the cross reacting material to move the antibody activity that causes the cross reaction. centrifuge down and only the unreacted target antibody will be left.
Describe the technique of affinity chromatography
Affinity Chromatography uses the target antigen to extract the target antibody and then uses high salt solution to wash off target antibody. This can be used to purify antibody or antigen.
Do monoclonal antibodies have cross-reactivity?
Yes but it is significantly lower than polyclonals
Describe the ability of flow cytometers and cell sorters to analyze and sort cells.
Flow cytometers can only analyze cells. Cell Sorters can both analyze and sort cells
describe the process of a Western Blot, what information does it give.
Proteins are separated by size electrophoretically, they are transferred to nitrocellulose, primary Ab is added, wash, secondary Ab is added, wash, and detect. Results are both quantitative and qualitative. possible to tell both the amount of antigen, its molecular weight, and different forms of the antigen you might be detecting.
