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Affinity Maturation by Phage Display

互联网

751
Antibodies are indispensable tools for research, diagnostics, and therapy. However, sometimes antibodies with the most favourable specificity profile lack sufficient affinity for a desired application. Here, we describe a method to increase the affinity of recombinant scFv antibody fragments based on random mutagenesis and phage display under stringent conditions. Random mutations are inserted by performing several rounds of error-prone PCR. After construction of a mutated antibody gene library, affinity selection is performed by panning with washing conditions optimized for off-rate-dependent selection. Alternatively, panning in solution with competition can be used to enrich binders with improved binding properties.
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