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filingDate 2002-03-28-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_9d7ee5edb861c4e22ef010557489b704
http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_137cb46c7e34fcd80e67319e66c15df9
publicationDate 2012-07-25-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber EP-2479280-A1
titleOfInvention Methods for the production of 3-O-deactivated-4'-monophosphoryl lipid A (3D-MLA)
abstract Herein is disclosed a method for producing lipopolysaccharide (LPS), comprising: (a) growing a culture of a deep rough mutant bacterial strain in a medium; (b) maintaining the culture in stationary phase for at least about 5 hr; (c) harvesting cells from the culture; and (d) extracting LPS from the cells. The method allows for the production of an LPS which can be used to produce a 3-0-deacylated monophosphoryl lipid A (3D-MLA) having at least about 20 mol% of the hexaacyl congener group. Also herein is disclosed a method of extracting lipopolysaccharide (LPS) from a culture of deep rough mutant bacterial strain cells, comprising: (a) extracting the cells with a solution consisting essentially of at least about 75 wt% of an aliphatic alcohol having from 1 to 4 carbon atoms and the balance water, thereby producing cells with reduced phospholipid content; and (b) extracting the cells with reduced phospholipid content with a solution comprising chloroform and methanol, thereby yielding a solution of LPS in chloroform and methanol (mM). This method provides LPS solutions in CM that have reduced phospholipid content and are produced by relatively simple and inexpensive process steps.
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