Cell-free expression of the outer membrane protein OprF of $Pseudomonas\ aeruginosa$ for vaccine purposes

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Mayeux, Géraldine | Gayet, Landry | Liguori, Lavinia | Odier, Marine | Martin, Donald | Cortès, Sandra | Schaack, Béatrice | Lenormand, Jean-Luc

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International audience. $Pseudomonas\ aeruginosa$ is the second-leading cause of nosocomial infections and pneumonia in hospitals. Because of its extraordinary capacity for developing resistance to antibiotics, treating infections by Pseudomonas is becoming a challenge, lengthening hospital stays, and increasing medical costs and mortality. The outer membrane protein OprF is a well-conserved and immunogenic porin playing an important role in quorum sensing and in biofilm formation. Here, we used a bacterial cell-free expression system to reconstitute OprF under its native forms in liposomes and we demonstrated that the resulting OprF proteoliposomes can be used as a fully functional recombinant vaccine against $P.\ aeruginosa$ . Remarkably, we showed that our system promotes the folding of OprF into its active open oligomerized state as well as the formation of mega-pores. Our approach thus represents an easy and efficient way for producing bacterial membrane antigens exposing native epitopes for vaccine purposes.

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