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Team:Potsdam Bioware/Project/Part Phage Display
From 2012.igem.org
Phage Display Test
Introduction
In order to characterize the AID and its potential to mutate antibody fragments, phage display is used. This powerful selection tool tests for proteins or peptides from a large recombinant library with specific binding abilities for a defined target. A gene of interest is introduced into the bacteriophage genome and will be presented on the surface of the phage allowing the selection for the phenotype closely coupled to the genotype.
We are planning to simultaneously transform E.coli cells with an arabinose inducible AID enzyme and a single chain antibody fragment EGFR-C on the phagemid vector pAK100. By allowing the enzyme to mutate the antibody fragment over a certain time we are generating a diverse library of antibody sequences. After the infection of E.coli cells with the helper phage, bacteriophages are produced and the mutated antibody sequence is randomly incorporated into their genome. Subsequently, E.coli cells are infected with the phages containing mutated antibody fragments. To observe the mutation rate of the antibody fragment, after two cycles of infection and phage production in presence of AID, multiple mutated phagemids are sequenced.
