US2007212758A1PendingUtilityA1

Methods for the production of 3-o-deactivated-4'-monophosphoryl lipid a (3d-mla)

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Assignee: CONXA CORPPriority: Mar 30, 2001Filed: Jan 9, 2007Published: Sep 13, 2007
Est. expiryMar 30, 2021(expired)· nominal 20-yr term from priority
Inventors:Kent Raymond MyersD. Snyder
C12P 19/04A61K 2039/55572C07H 13/06A61P 37/04C12N 1/20
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Claims

Abstract

Herein is disclosed a method for producing lipopolysaccharide (LPS), comprising: (a) growing a culture of 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-O-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 (CM). This method provides LPS solutions in CM that have reduced phospholipid content and are produced by relatively simple and inexpensive process steps.

Claims

exact text as granted — not AI-modified
1 - 25 . (canceled)  
   
   
       26 . A composition of 3D-MLA comprising at least about 20 mol % of hexaacyl congener.  
   
   
       27 . A composition according to  claim 26  comprising between 20 and 50 mol % of hexaacyl congener  
   
   
       28 . A composition according to  claim 26  comprising at least 30 mol % of hexaacyl congener.  
   
   
       29 . A composition according to  claim 26  comprising 21.5% hexaacyl congener.  
   
   
       30 . A composition of LPS comprising at least about 20 mol % of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       31 . A composition according to  claim 30  comprising at least 30 mol % of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       32 . A composition according to  claim 30  comprising 36% of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       33 . A composition of MLA comprising at least about 20 mol % of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       34 . A composition according to  claim 33  comprising at least 30 mol % of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       35 . A composition according to  claim 33  comprising 32.7% of a combination of heptaacyl congener and 3-O-deacylated hexaacyl congener.  
   
   
       36 . A composition according to  claim 26  wherein said 3D-MLA is extracted from a deep rough mutant strain of a gram-negative bacterium.  
   
   
       37 . A composition according to  claim 30  wherein said LPS is extracted from a deep rough mutant strain of a gram-negative bacterium.  
   
   
       38 . A composition according to  claim 33  wherein said MLA is extracted from a deep rough mutant strain of a gram-negative bacterium.  
   
   
       39 . A composition according to  claim 36  wherein said 3D-MLA is extracted from a  Salmonella  bacteria.  
   
   
       40 . A composition according to claims  30  wherein said LPS is extracted from a  Salmonella  bacteria.  
   
   
       41 . A composition according to claims  33  wherein said 3D-MLA is extracted from a  Salmonella  bacteria.  
   
   
       42 . A composition according to  claim 34  wherein said bacterium is  Salmonella Minnesota.    
   
   
       43 . A composition according to  claim 35  wherein said bacterium is strain  Salmonella Minnesota  R595.  
   
   
       44 . A plurality of compositions of 3D-MLA having a consistent hexaacyl content.  
   
   
       45 . A plurality of compositions according to  claim 44  wherein said hexaacyl content is consistently at least about 20 mol %.  
   
   
       46 . A plurality of compositions according to  claim 44  wherein said hexaacyl content is consistently between 20 mol % and 50 mol %.  
   
   
       47 . A plurality of compositions according to  claim 44  wherein said hexaacyl content is consistently at least about 50mol %.

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