phage display technology - creative-biolabs.com · phage display library eluting amplify...
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M13 coat protein 3 (g3p), present on average in 5 copies at the tip of the M13 phage particle, is often the first choice for phage display fusions because of its tolerance for large insertions.
Filamentous phage M13 is the mostly commonly used for antibody display.
Directly insertion makes the phage plasmid too large to transfection and amplification.
A phagemid and helper phage system is commonly used for antibody display.
Monovalent display is preferred due to the demanding of high affinity monoclonal Abs.
g1p: Morphogenesisg4p,
M13T7
T4λ
G7P G9P
G8P
ssDNA M136.4Kb
G9PG3P
g2p: RF replicationEndonuclease
g10pg5p: ssDNA bindingg7p: Minor capsidg8p: Major capsid
g3p: Absorbtion protein
g6p: Minor capsid
Phage: structure and genome
Phagemid system for antibody display
pHAL30
Ab Heavy Chain
bla
g3pLac Pr.Stuffer fragment
Ab Light Chain
Phagemid E. coli
Helper Phage
Packaged Phage with antibody displayedon surface
Antibody
G3P
Ab DNA library
Drug Discovery
Phage plasmid
Recombinantphagemid
2-3 Rounds
High affinity mAb
Weak affinity Abs
Affinity screening
Binding and Wash
Phage display library
Eluting
Amplify
Diagnostics
AntibodyProduction
1
2
Phage display library and screening3
Solid surface immobilized with antigen is used for antibody phage screening.
Antibody phage particles that bind weakly to the antigen and the vast excess of non-binding antibody phages are removed by stringent washing. Specifi-cally binding antibody phage are eluted and used for infection of for the next round screening.
2-3 round of screening are necessary for monoclonal antibody with high enough affinity.
Immune, naïve or synthetic library constructionPre-made library screeningLarge library capacityHigh affinity from 10-7 to 10-9
Various phage display systems (M13, T4, T7)Tailored biopanning strategies
Phage display library construc-tionPhage display library screeningSpecific antibody discoveryPeptidomic discovery
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PHAGE DISPLAY TECHNOLOGY
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