RT Journal Article SR Electronic T1 Cell-free expression of the outer membrane protein OprF of Pseudomonas aeruginosa for vaccine purposes JF Life Science Alliance JO Life Sci. Alliance FD Life Science Alliance LLC SP e202000958 DO 10.26508/lsa.202000958 VO 4 IS 6 A1 Géraldine Mayeux A1 Landry Gayet A1 Lavinia Liguori A1 Marine Odier A1 Donald K Martin A1 Sandra Cortès A1 Béatrice Schaack A1 Jean-Luc Lenormand YR 2021 UL https://www.life-science-alliance.org/content/4/6/e202000958.abstract AB 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.