Bacteria-derived vesicles and uses thereof

A method for generating non-naturally occurring artificial outer membrane vesicles (aOMVs) from a gram-negative bacterium, the method comprising:

• a) disrupting a spheroplast generated from the gram-negative bacterium to generate vesicles comprising outer membrane and vesicles comprising inner membrane, optionally wherein disrupting comprises applying shear force to the spheroplast or acoustic energy to the spheroplast;
• b) exposing the vesicles to an ionic surfactant to disrupt vesicles comprising inner membrane and to an alkaline pH to open the vesicles comprising outer membrane thereby generating outer membrane sheets;
• c) purifying the outer membrane sheets;
• and
• d) applying energy to the purified outer membrane sheets sufficient to convert the outer membrane sheets into aOMVs, thereby generating the-non-naturally occurring aOMVs.

A method for generating non-naturally occurring artificial outer membrane vesicles (aOMVs) from a gram-negative bacterium, the method comprising:

• a) disrupting a spheroplast generated from the gram-negative bacterium to generate vesicles comprising outer membrane and vesicles comprising inner membrane, optionally wherein disrupting comprises applying shear force to the spheroplast or acoustic energy to the spheroplast;
• b) exposing the vesicles to an ionic surfactant to disrupt vesicles comprising inner membrane and to an alkaline pH to open the vesicles comprising outer membrane thereby generating outer membrane sheets;
• c) purifying the outer membrane sheets;
• and
• d) in the presence of a STING agonist, applying energy to the purified outer membrane sheets sufficient to convert the outer membrane sheets into aOMVs, thereby generating the non-naturally occurring aOMVs comprising the STING agonist.