L5_Immunoassays

Question 1

Define Avidity

Answer 1

The strength of binding of multivalent antiserum to multivalent antigen

Question 2

what are the two main techniques used to eliminate cross-reactivity

Answer 2

absorption and affinity chromatography

Question 3

Describe Monoclonal Antibody Affinity, Avidity and cross reactivity compared to polyclonal

Answer 3

They have lower cross reactivity, lower affinity and little avidity.

Question 4

What is a myeloma cell?

Answer 4

A tumor of a plasma cell that no longer creates antibodies by has the capability to do so.

Question 5

Describe the process of making a monoclonal antibody

Answer 5

1. Immunize an animal
2. Isolate spleen cells form immunized animal
3. Fuse spleen cells to plasmacytoma tumor cells (myeloma)
4. Select for only those cells that are hybrids
5. Clone hybridomas so each single cell grows up independently
6. selelct the individual clone with the specificity you are interested in

Question 6

What will a drug name end with if it is a monoclonal antibody

Answer 6

mab

Question 7

What is the function of framework residues?

Answer 7

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.

Question 8

Describe the technique of absorption

Answer 8

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.

Question 9

Describe the technique of affinity chromatography

Answer 9

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.

Question 10

Do monoclonal antibodies have cross-reactivity?

Answer 10

Yes but it is significantly lower than polyclonals

Question 11

Describe the ability of flow cytometers and cell sorters to analyze and sort cells.

Answer 11

Flow cytometers can only analyze cells. Cell Sorters can both analyze and sort cells

Question 12

describe the process of a Western Blot, what information does it give.

Answer 12

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.