{[ promptMessage ]}

Bookmark it

{[ promptMessage ]}

Bi1_2009_Lecture23_full

Bi1_2009_Lecture23_full - Putting biology to work for you...

Info iconThis preview shows pages 1–6. Sign up to view the full content.

View Full Document Right Arrow Icon
Putting biology to work for you: In vitro (directed) evolution and other techniques
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Phage display • Make a combinatorial library of genes of interest • Put genes into a vector so that each gene product is expressed on the surface of a bacteriophage – Function encoded by each gene is on surface of phage – Gene is inside phage • Take collection of phage and select those with desired properties (e.g., binding to something) • Similar methods: yeast display, bacterial display, ribosome display See animation of phage display here: http://www.dyax.com/discovery/phagedisplay.html
Background image of page 2
Figure A-15 In vitro selection to produce human monoclonal antibodies or increase affinity of existing monoclonal antibody Generate library of heavy and light chain variable regions using spleen DNA. Or introduce random mutations into variable regions genes of a specific antibody. Clone into a phage so that each phage expresses one V H -V L surface fusion protein. Multiply phage display library in bacteria, bind phage to surface coated with antigen. Wash away unbound phage. Repeat procedure (multiply recovered phage, bind to antigen, wash away unbound phage) for several cycles. Recover specific high-affinity antigen binding V H -V L regions.
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Phage Display schematic 1 2 3 4 5 6 7 8
Background image of page 4
Phage display to select for sequence-specific DNA binding proteins • Zinc finger proteins are modular.
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Image of page 6
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}