L13

Question 1

what is a hybridoma

Answer 1

has AB producing genes from B cell and the immortal properties fo a tumout cell

Question 2

process of making MAbs in vitrro

Answer 2

Mice are immunised with an antigen – a couple of times
Spleen cells are removed
Contain the antigen-specific B cells
These are mortal i.e. finite life span
Splenocytes “fused” in vitro with myeloma cell line
1 cell is plated and as these proliferate will have monoclonal antibodies

Question 3

what growth medium are they cultured in

Answer 3

contains HAT medium
(hypoxanthine, aminopterin and thymidine)
so only fused cells will survive

Question 4

advantages of MAbs

Answer 4

High specificity
Low cross-reactivity
Standardised worldwide
Unlimited supply

Question 5

issues MABs can cause

Answer 5

trigger immune responses
NOT access the antigen sites needed
Cause off target effects - maybe epitope is shared

Question 6

solutions to some of the problems

Answer 6

make in in plants/insects instead of mouse
Fab only
delivery systems to target cells

Question 7

what are MAbs used for

Answer 7

affinity chromatography
magnetic separation if cell populations
haemaagglutination

can tag the antibody
- ELISA
- elispot
- immunofluoresence

Question 8

what is an ELISpot

Answer 8

used in situ
technique similar to elisa
but the substrate makes a precipitating product not just a colour change = visible spots on the surface
can see frq of individual cells