Simplified low-cost production of O-antigen from Salmonella Typhimurium Generalized Modules for Membrane Antigens (GMMA)

• Published in: Journal of biotechnology Vol. 198; pp. 46 - 52
• Main Authors: Meloni, Eleonora, Colucci, Anna Maria, Micoli, Francesca, Sollai, Luigi, Gavini, Massimiliano, Saul, Allan, Di Cioccio, Vito, MacLennan, Calman A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 20.03.2015
• Subjects: Antigens; Bacteria; GMMA; Hydrolysis; Lipopolysaccharide; Lipopolysaccharides - chemistry; Lipopolysaccharides - metabolism; Membranes; Membranes - chemistry; Membranes - metabolism; Modules; O Antigens - chemistry; O Antigens - isolation & purification; O Antigens - metabolism; O-antigen; Polysaccharides; Purification; Salmonella; Salmonella typhimurium; Salmonella typhimurium - chemistry; Salmonella typhimurium - metabolism; Vaccine; Vaccines; Vaccines - chemistry; Salmonella; HMW; TFF; MMW; LMW; Lipopolysaccharide; O-antigen; Vaccine; GMMA; CDM; LPS
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2015.01.020

•GMMA are a potential source of outer membrane components for vaccine manufacture.•Large quantities of GMMA are released during the fermentation of ΔtolR Salmonella.•GMMA are enriched in LPS, but contain low quantities of nucleic acids and proteins.•Purification of O-antigen from GMMA requires fewer steps than from whole bacteria.•GMMA as starting material for vaccine production has potential to lower cost-of-goods. 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.