Case unit 7

Question 1

what is screening

Answer 1

find something and then work out how it works

Question 2

what is an assay

Answer 2

you know what you are looking for and want and then find something that will do it

Question 3

advantages of screening

Answer 3

quick
you dont need previous knowledge
already present so you dont need to make anything
cheap

Question 4

why do PCR

Answer 4

intorduce variation
cause mutations to occur
allows you to select the best ones

Question 5

what is a dependant varaible

Answer 5

looking to see the change in response to the independant

Question 6

what is a independant varaible

Answer 6

something that changes and causes an effect on the dependant

Question 7

what is a confounding varaiable

Answer 7

changing varaiable that could influence on dependant

Question 8

phage display

Answer 8

Use in vitro evolution
Directed evolution
High selective pressure
Like breeding dogs

Question 9

steps of invtro evolution - therauptic antibodies

Answer 9

Fuse the cDNA for a Fab to a phage coat protein
Genetically engineer a library of bacteriophages (M13) with many different cDNA sequences (>109)
Expose phages to an immobilised target molecule
Use high stringency washes to leave the strongest binders attached to the ligand
Elute strong binders
Use strong binders to infect bacteria
Natural mutations will occur when the bacteria are grown up and new phages produced.
New variants on a theme are produced
Repeat the process
Increase wash stringency
Third generation phages produced
Repeat cycle till you have a super-high affinity binder