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Matt--Phage display

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  • 1. Phage Display and its Applications Matt Brown Human Genetics Dr. Nancy Bachman
  • 2. Basics
    • 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
  • 3. Initial Steps In Phage Display
    • 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
  • 4. Common Phages used in Display
    • 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
  • 5. Phage Display Screening
    • 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
  • 6. Anti-Body Engineering
    • 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
  • 7. Isolating Selected Antigens
    • 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
  • 8.
    • Once these Phages are isolated and recovered they can be used to infect bacteria which will create a particle similar to a monoclonal antibody
    Isolating Selected Antigens Cont’d
  • 9. Conclusion
    • 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
  • 10. References
    • Strachan Tom, Human Molecular Genetics 3. 2004 Garland Publishing; pp-150-151
    • http://www.antibodystation.com/phage-display-antibody-production/
    • Journal Of Virology 2001 pp. 75
    • Nature 2000 pp. 45