Display library process
Disclosed are methods for identifying desired members from a display libraries, including bacteriophage display libraries. Display library members can be amplified in the presence of a target compound so that cycles of selection can be rapidly completed.
1. A method of selecting phage that encode a target binding protein from a plurality of phage particles, the method comprising:
a) contacting a plurality of diverse phage particles with a first target to form phage-target complexes, wherein each phage of the plurality displays a heterologous protein component on its surface and each phage includes a nucleic acid encoding the heterologous protein component, and wherein each phage-target complex comprises a phage from the plurality, the heterologous protein displayed on which binds the first target;
b) separating the phage-target complexes from unbound phage particles;
c) contacting host cells with the separated phage-target complexes in a suitable growth medium so that the host cells are infected by the phage particles of the phage-target complexes to yield a first population of infected cells;
d) incubating the first population of infected cells in the presence of a second target under conditions allowing for replication of the phage particles, thereby forming phage-target complexes that include replicate phage particles that bind to the second target;
e) separating replicate phage particles that do not bind to the second target from the replicate phage-target complexes; and
f) contacting host cells with the replicate phage target complexes so that host cells are infected with the replicate phage particles of the complexes to yield a second population of infected cells, wherein the second population of infected cells produce phage particles that encode heterologous proteins binding to both the first target and the second target;
wherein the first target and the second target are different.
2. The method of claim 1 , wherein the first target is immobilized to a support prior to step (a).
3. The method of claim 1 , wherein the first target is immobilized to a support during step (a).
4. The method of claim 1 , wherein the second target is immobilized to a support prior to step (d).
5. The method of claim 1 , wherein the second target is immobilized during step (d).