US EPA

7439413 

Journal Article 

Whole-Bacterium Ribosome Display Selection for Isolation of Antibacterial Affitins 

Béhar, G; Kambarev, S; Jazat, J; Mouratou, B; Pecorari, F; , 

2020 

153-171 

10.1007/978-1-0716-0720-6_9 

http://link.springer.com/10.1007/978-1-0716-0720-6_9
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Classically, in order to perform ribosome display selections, a randomized gene library encoding potential affinity proteins is incubated with a pure target of interest that is either immobilized on a solid surface such as microtiter plates and paramagnetic/agarose beads or used in solution in a way that allows the capture of specific binders. The success of these selections mainly depends on the availability and conformation of the target used. In some cases, the most relevant target can be unknown, or it can be impossible to get it in its native state. Here, we describe a method to perform ribosome display selections against targets at the surface of live bacterial cells. The protocol explains the preparation of bacteria as selection target, panning with libraries, elution, and screenings procedures. We exemplify this strategy with the selection of Sac7d-based Affitins against bacteria as target.