Phage Display and Selections on Biotinylated Antigens
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In vitro selections performed by phage display are often performed against a recombinant antigen adsorbed on a solid surface. Passive adsorption of the antigen involves its partial denaturation, which can lead to binders able to strongly recognize the adsorbed antigen but displaying a poor affinity for the native antigen. As an alternative, the use of in vitro or in vivo biotinyled antigens avoids such issues, while allowing a precise control of the interaction between the phage particle and the antigen that takes place in solution. The whole complex can be retrieved by a short incubation over a streptavidin-coated solid phase. This chapter provides detailed protocols to perform in vitro or in vivo biotinylation of the antigen and phage display selection and screenings on biotinylated antigens.
KeywordsPhage Particle Phage Antibody Inhibition ELISA Specific Phage Positive Plate
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