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Phage Display and its Applications

Phage Display and its Applications. Matt Brown Human Genetics Dr. Nancy Bachman. 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. Basics.

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Phage Display and its Applications

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  1. Phage Display and its Applications Matt Brown Human Genetics Dr. Nancy Bachman

  2. 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 Basics

  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. Isolating Selected Antigens Cont’d • Once these Phages are isolated and recovered they can be used to infect bacteria which will create a particle similar to a monoclonal antibody

  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

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