Methods and culture media for production, quantification and isolation of bacteriophages

Abstract

The present invention is directed to a method for producing bacteriophages infecting a bacterial host such asorsp, the method comprising the steps of adding the bacterium to a sterile culture medium supplemented with mucin or another mucus component, incubating said culture medium, thereby obtaining a bacterial culture, optionally collecting the supernatant containing floating (planktonic) bacterial cells from said culture and transferring the supernatant to a new container in order to discard most of the biofilm and adding a bacteriophage infecting the bacterium to said bacterial culture obtained from a previous step, and incubating the culture until the phage yield increases or peaks. The method can also be used for preparing modified enrichment cultures for optimized phage isolation. Addition of mucin to soft-agar culture medium is proposed as an optimized technique for viral titration. This invention also covers all relevant steps in setting up a phage therapy product: phage production, phage quantification and phage isolation.