Phage_display

Question 1

what is phage display technology

Answer 1

is in vitro selection method that allows isolation of antigen specific antibody from a large number of collected molecule

Question 2

what is the role of phase display technique in antibody production

Answer 2

it allows in vitro selection and production of recombinant monoclonal antibody of high specificity and affinity

Question 3

what is the Comparison of innate immune response/ in vivo and Phage display

Answer 3

page 4

Question 4

where can recombinant ab bee expressed on in phage display

Answer 4

ab fragment can be expressed in bacteria

Question 5

where can we get antibody library diversity

Answer 5

• gene encoded ab can be isolated from B cells
• gene encoded ab can be isolated from CDR

gene can be from human or animal

Question 6

how to get Antibody libraries

Answer 6

donor can be human or animal
then gene encoding are isolated in B cells form either immunized or non-immune subject
PCR VH and VL are assembled
scFv antibody fragment is encoded with VH-VL

Question 7

how to find what you want (ab) in different variants

Answer 7

by using phage display because phage displays ab fragment on its surface

Question 8

what is a phage

Answer 8

is a virus that infects bacteria

Question 9

what is filamentous phage

Answer 9

is long and thin particle that contain circular ssDNA

Question 10

can any protein be displayed on phage

Answer 10

no, like full length antibodies can be displayed on phage

Question 11

How do you get the antibody fragment on the surface of the phage?

Answer 11

protein III display antibody on its surface, how:
1. E.cole pilus
2. transformation with protein encoding phagemid vector
3. ampicillin resistant transformants : this help us to select E.coli cells that has been transformed
4. infection with helper phage: help in supply of structural protein required to generate complete phage. undergoes induction
5. production of phage displaying fuse protein

Question 12

what is phagemid

Answer 12

is a hybrids of phage and plasmid vector

Question 13

what is the mechanism of Phage production of desired property

Answer 13

1. immobilized antigen
2. phage library: here is binding of phage to antigen
3. washing unbound phage
4. elute bind phage and infected E.coli
5. amplify (increase in volume) binding phage
6. repeat step 1-5 till you reach on sufficient enrichment
7. screen for desired properties: to identify variants with desired properties
8. mutate gene encoding binder
9. create a new phage library and phage stock

Question 14

what is mutation of gene

Answer 14

is changing DNA sequence

Question 15

what ate phage and selection technologies

Answer 15

• phase display
  -bacterial display
• yeast display
• ribosomal display

Question 16

what is the importance of Display and selection technologies

Answer 16

allows display and selection of variant number of population and link between functional and genetic information

Question 17

how to analyze selected binders

Answer 17

• first wash unbound phage and amplify it in E.coli and repeat.
• infect E.coli and pick single colony into a well
• have to be monoclonal
• the analyze assay for binding by immunoassay

Question 18

what is the advantage of phage display

Answer 18

• increase binding properties of initial antibody selection
• allows generation of antibodies for human origin
• allow better selection condition
• give better efficiency than hybridoma method

Question 19

application of phage display

Answer 19

• generation of monoclonal antibodies
• improving existing monoclonal antibody
• studying protein-protein/ antigen-antibody interaction
• studying antibody response

Question 20

hry