The Novartis
Vaccines Institute for Global Health is developing
vaccines using outer membrane particles, known as Generalized Modules for Membrane
Antigens (
GMMA). These are
blebs of outer membrane and periplasm, shed from the surface of living Gram-negative bacteria following the targeted deletion of
proteins involved in maintaining the integrity of the inner and outer membranes. The current study investigates the use of
GMMA as starting material for extraction of membrane components, focusing on the
O-antigen polysaccharide portion of
lipopolysaccharide from Salmonella Typhimurium. We show that the amount of
O-antigen extracted from
GMMA by
acid hydrolysis is comparable to the quantity extracted from whole wild type bacteria, but with less
protein and
DNA contaminants. Compared to conventional purification,
GMMA enabled a reduction in the number of purification steps required to obtain the
O-antigen polysaccharide with the same purity. Purification processes from
GMMA and bacteria were characterised by similar final yields. Use of
GMMA as starting material provides the possibility to simplify the purification process of
O-antigen, with a consequent decrease in manufacturing costs of
O-antigen-based glyconjugate
vaccines against Salmonella strains and potentially other Gram-negative bacteria.