Phage display Flashcards
Phage display
a technique in which antibody fragments are expressed as a fusion coat protein on bacteriophage
fused proteins (antibody fragments)
are displayed on the surface of the vision, while the DNA encoding the fusion resided within the virion
DNA encoding the antibody (fusion)
is found within the virion
what sort of phage is used
a filamentous phage structyre
name an example of a filamentous phage
M13 phage
how long in length
200nm
what sort of genome
ssDNA
target antibody to be expressed
on the outside of the vision
process of making a phage display library
1) B cell source (immunised animal)
2) production of mRNA
3) production of cDNA from mRNA
4) PCR to amplify each the the V-gene facilities ( VH and VL)
5) clone as scFv onto PIII of phage
scFv =
Vl and Vh of an immunogoblin linked by a peptide spacer
what allow expression of scFv on the surface of phages
fusion proteins between scFV and PIII minor coat proteins allow expression
where are PIII minor coat proteins found
on the end of the phage
Fab fragments of antibodies can be expressed on the
surface of a phage as fusion proteins with PIII
Fab fragments of antibodies can also be expressed on the surface of phages as fusion proteins with
pVIII
where is pVIII expressed
along the length of the phage particle