抗体噬菌体展示技术

▪ Capable of generating antibodies against almost any
desired antigen, including highly conserved or
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self-antigens, conformational variants, low
immunogenic antigens, and also toxic components,
beads, polystryrene surfaces, or on columns, or is used in solution as
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Advantages of Phage Display for Recombinant
Antibody Selection
▪ Cheaper to produce recombinant antibodies using
bacteria, rather than mammalian cell line.
▪ Easier to maintain and grow bacterial cultures for
recombinant antibody production.
▪ Bypass immunization in antibody selection.
▪ Promoter
▪ Signal peptides: phage protein
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translocation, crucial for display level
Introduction of Phage Display Technology
▪ Nonlytic filamentous phage is the
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Introduction of Phage Display Technology
The Ff bacteriophage structure
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Introduction of Phage Display Technology
The scheme of phagemid vector
▪ IG region: intergenic region, usually
most often used for phage display, primarily the M13 and Fd strains.
▪ Proteins to be selected are infused
to all five coat proteins, with pIII and pVIII most commonly used.
▪ pIII protein is essential for
infection of bacteria
▪ Helper phage: wild-type pIII helper
phage and special helper phage
▪ Antigen immobilized on magnetic
Antibody Phage Display
Meiling Xiong 20180629
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Contents
▪ Introduction of Ab phage Display Technology ▪ Ab Formats for Phage Display ▪ Ab Libraries Construction ▪ Phage Ab Selection Methods & Strategies ▪ Phage Ab Screening Applications ▪ In vitro Affinity Maturation ▪ Expression & Purification of Phage Ab Fragments
which is not possible by in vivo immunization of
than 5 copies,) PVIII (more than 5 copies,
decreased length)
▪ Amber codon TAG: supE strains (glutamic
acid codon), non-suppressor strains (stop
codon)
▪ Protease cleavage site
▪ Easy isolation and expression of the cloned gene in
a bacterial host.
▪ Excellent potential to further improve binding
properties of the selected antibody by protein engineering techniques.
▪ Bypass the use of animal cells for production of
antibodies.
▪ Producing the combinatorial library (ideally with
108 to 109 members) of functional antibodies to generate a larger repertoire of antibodies than those available through conventional hybridoma technology.
▪ Restriction enzyme recognition sites:
useful for DNA recombination and gene
manipulation; multiple cloning sites (MCS)
▪ Coat protein: PIII (larger protein, less
contains the packing sequence and replication origin of minus and plus strands
▪ Molecular tag: to facilitate library
screening and for protein analysis