Rapid dehydration of proteins

Abstract

The present invention relates to protein-coated micro-crystals and their method of preparation. The protein-coated micro-crystals may find particular application in preparing enzymes for use as biocatalysts; preparation of therapeutic proteins for use in pharmaceutical formulations; production of cleansing agents comprising enzymes; production of paints, varnishes, coatings, films and the like comprising proteins which impart protective and/or antifouling properties; production of films, polymers, inks, coatings, electrodes and/or optical materials comprising proteins for diagnostic kits and/or biosensor applications; use of proteins for studies of molecular recognition, molecular binding and inhibitor binding in non-aqueous media; and preparation of protein based food additives.

1 . Water soluble particles comprising micro-crystals of less than 50 μm, said micro-crystals comprising a crystalline coprecipitant core with a dehydrated biological macromolecule coated thereon; 
 wherein the coprecipitant core consists of one of the following:    inorganic salts,    sugars, polyols, and derivatives thereof with a molecular weight of less than 10,000 Da;    amino-acids;    acid-base buffers;    zwitterionic compounds;    organic salts;    compounds containing multiple basic groups;    compounds containing multiple acidic groups;    bile salts; and,    water soluble dyes.    
     
     
         2 . Water soluble particles according to  claim 1  wherein the dehydrated biological macromolecule is selected from peptides, polypeptides, proteins and nucleic acid.  
     
     
         3 . Water soluble particles according to  claim 1  having a diameter less than 10 μm.  
     
     
         4 . Water soluble particles according to  claim 1  wherein the coprecipitant is trehalose.  
     
     
         5 . Water soluble particles according to  claim 1  wherein the coprecipitant is an amino acid selected from the group consisting of glycine and arginine.  
     
     
         6 . A pharmaceutical formulation comprising particles according to  claim 1  and a suitable carrier therefor.  
     
     
         7 . A medical device comprising particles according to  claim 1  associated therewith.  
     
     
         8 . Water soluble particles according to  claim 1  wherein said coprecipitant core is a non-polymeric core.  
     
     
         9 . Particles according to  claim 1  for use in therapy.  
     
     
         10 . A biocatalyst preparation comprising particles according to  claim 1  associated therewith.  
     
     
         11 . A cleansing agent comprising enzyme coated particles according to  claim 1 .  
     
     
         12 . A protective or antifouling agent comprising particles according to  claim 1  in association with paint, varnish, coatings or films.  
     
     
         13 . Films, polymers, inks, coatings, electrodes and optical materials for diagnostic kits or biosensor applications, comprising particles according to  claim 1 .  
     
     
         14 . A method for studying molecular recognition, molecular binding, molecular imprinting or inhibitor binding in non-aqueous media, comprising using particles according to  claim 1 .  
     
     
         15 . A method for studying macromolecule structure and/or organisation by scanning probe microscopy, comprising using particles according to  claim 1 .  
     
     
         16 . A pharmaceutical formulation comprising biological macromolecule coated micro-crystals of less than 50 μm comprising a coprecipitant core with a dehydrated pharmaceutically active biological macromolecule coated thereon; 
 wherein the coprecipitant core consists of one of the following:    inorganic salts,    sugars, polysaccharides, polyols, and derivatives thereof with a molecular weight less than 10,000 Da;    amino-acids;    acid-base buffers;    zwitterionic compounds;    organic salts;    compounds containing multiple basic groups;    compounds containing multiple acidic groups;    bile salts; and    water soluble dyes;    and a suitable carrier therefor.    
     
     
         17 . The pharmaceutical formulation according to  claim 16  wherein the coprecipitant is trehalose.  
     
     
         18 . The pharmaceutical formulation according to  claim 16  wherein the coprecipitant is an amino acid selected from the group consisting of glycine and arginine.  
     
     
         19 . An inhalable pharmaceutical formulation comprising biological macromolecule coated micro-crystals comprising a coprecipitant core with a dehydrated pharmaceutically active biological macromolecule coated thereon; 
 where the coprecipitant core consists of one of the following:    inorganic salts,    sugars, polysaccharides, polyols, and derivatives thereof with a molecular weight less than 10,000 Da;    amino-acids;    acid-base buffers;    zwitterionic compounds;    organic salts;    compounds containing multiple basic groups;    compounds containing multiple acidic groups;    bile salts; and    water soluble dyes;    and a suitable carrier therefor.    
     
     
         20 . A liquid suspension comprising water soluble particles comprising micro-crystals of less than 50 μm, said micro-crystals comprising a crystalline coprecipitant core with a dehydrated biological macromolecule coated thereon; 
 wherein the coprecipitant core consists of one of the following:    inorganic salts,    sugars, polyols, and derivatives thereof with a molecular weight less than 10,000 Da;    amino-acids;    acid-base buffers;    zwitterionic compounds;    organic salts;    compounds containing multiple basic groups;    compounds containing multiple acidic groups;    bile salts; and    water soluble dyes.    
     
     
         21 . A method of purifying a biological macromolecule from additives or impurities comprising: 
 a) dissolving a coprecipitant in an aqueous solution comprising the biological macromolecule and additive or impurity wherein the coprecipitant is selected from the group consisting of inorganic salts; sugars, polyols, and derivatives thereof with a molecular weight less than 10,000 Da; amino-acids; acid base buffers; zwitterionic compounds; organic salts; compounds containing multiple basic groups; compounds containing multiple acidic groups; bile salts; and, water soluble dyes;    b) admixing the biological macromolecule/coprecipitant solution with an excess of a water miscible organic solvent or solvents, in which the additive or impurity is soluble, such that the coprecipitant and biological macromolecule immediately coprecipitate from solution forming a biological macromolecule coated micro-crystals of less than 50 μm.    
     
     
         22 . A method of purifying a biological macromolecule from additives or impurities wherein said method is used or isolating the biological macromolecule from the aqueous solution.