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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