Transcript Phage Display and its Applications

Phage Display and its
Applications
Matt Brown
Human Genetics
Dr. Nancy Bachman
Basics
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Used for cloning foreign
genes among other
applications
Proteins and peptides
are fused to the
Capsid(surface) of the
phage
The combination of the
phage and peptide is
known as a Fusion
Protein
Initial Steps In Phage Display
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Different sets of genes are
inserted into the genomes
of multiple phages
These separate phages will
only display one protein,
peptide, or antibody
Collections of these phages
can comprise Libraries
These Libraries are exposed
to selected targets and only
some phages will interact
with targets
Common Phages used in
Display
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3 types of common phages used in
phage display are the M13, F1 , FD
Virions take up a small amount of area
Through using multiple Virions
polypeptide libraries can be created,
and each phage displays a random
peptide
Phage Display Screening
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Polypeptides of interest
can be screened using
selection techniques
The target
protein/peptide can be
immobilized using
magnetic beads
With the advance of
DNA sequence
recognition these
selected sequences can
be identified easily
Anti-Body Engineering
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By taking gene segment of antigens of
antibodies and fusing them to the protein
coat of phages, these phages will now
express the anti-body in a fusion protein
Phage Display Libraries of antigens can be
created to create anti-body phage display
libraries
Isolating Selected Antigens
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The same methods are used to isolate these
selected antigens as a regular polypeptide
phage.
One example of this besides using magnetic
beads is affinity chromatography
(separation of biochemical solutions)
If a solution known to contain antibodies
against a specific antigen, then the phage
expressing specific antigen can be isolated by
selecting the phage in the library for binding
to that antigen. it will produce antibodies
against the fusion-protein
Isolating Selected Antigens
Cont’d
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Once these Phages are isolated and
recovered they can be used to infect
bacteria which will create a particle
similar to a monoclonal antibody
Conclusion
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Proteins Isolated through phage display
will have a specific interactions with its
target
Whether this target is a antigen,
protein, or disease it allows a quick
effective treatment for it
References
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4.
Strachan Tom, Human Molecular
Genetics 3. 2004 Garland Publishing;
pp-150-151
http://www.antibodystation.com/phag
e-display-antibody-production/
Journal Of Virology 2001 pp. 75
Nature 2000 pp. 45