title: Simplified low-cost production of O-antigen from Salmonella Typhimurium Generalized Modules for Membrane Antigens (GMMA).
abstract: 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.
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Eleonora
authors: E
authors: Meloni
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Anna Maria
authors: AM
authors: Colucci
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Francesca
authors: F
authors: Micoli
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Luigi
authors: L
authors: Sollai
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Massimiliano
authors: M
authors: Gavini
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Allan
authors: A
authors: Saul
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.
authors: Vito
authors: V
authors: Di Cioccio
authors: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy. Electronic address: cm15@sanger.ac.uk.
authors: Calman A
authors: CA
authors: MacLennan
mesh_terms: D006868:Hydrolysis; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008566:Membranes / Q000737:chemistry / Q000378:metabolism*; D019081:O Antigens / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000737:chemistry* / Q000378:metabolism*; D014612:Vaccines / Q000737:chemistry
chemical_list: D008070:Lipopolysaccharides; D019081:O Antigens; D014612:Vaccines
keywords: GMMA; Lipopolysaccharide; O-antigen; Salmonella; Vaccine