Elution of biomolecules from multi-modal resins using mes and mops as mobile phase modifiers

Abstract

The present invention relates to methods for purifying or enriching a biomolecule using multimodal resins and an elution buffer containing a Good's buffer.

1 . A method for purifying or enriching a biomolecule, said method comprising the steps:
 (a) providing an aqueous solution containing said biomolecule;   (b) contacting said aqueous solution with a multimodal resin;   (c) optionally washing the multimodal resin with an aqueous washing buffer;   (d) eluting the biomolecule from the multimodal resin by an aqueous buffer solution,
 wherein said aqueous buffer solution comprises one or more of: 
 (i) at least 15 mM of a Good's buffer, 
 (ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine; 
 (iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and 
 (iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol; and 
   (e) collecting biomolecule containing fractions, thereby obtaining the biomolecule in purified or enriched form.   
     
     
         2 . The method according to  claim 1 , wherein the aqueous buffer solution comprises 15 mM to 2.5 M Good's buffer. 
     
     
         3 . The method according to  claim 1  or  claim 2 , wherein the Good's buffer is MES, MOPS, or Tris. 
     
     
         4 . The method according to  claim 1  or  claim 2 , wherein the Good's buffer is MES. 
     
     
         5 . The method according to  claim 1  or  claim 2 , wherein the Good's buffer is MOPS. 
     
     
         6 . The method according to any one of  claims 1  to  5 , wherein the biomolecule is a protein. 
     
     
         7 . The method according to  claim 6 , wherein the protein is an enzyme. 
     
     
         8 . The method according to  claim 7 , wherein the enzyme is a glycosidase. 
     
     
         9 . The method according to  claim 8 , wherein the glycosidase is an alpha-galactosidase. 
     
     
         10 . The method according to  claim 9 , wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta. 
     
     
         11 . The method according to any one of  claims 1  to  10 , wherein the multimodal resin is selected from the group consisting of Capto™ Adhere, Capto™ MMC, Pall PPA HyperCel™, Pall HEA HyperCel™, Pall MEP HyperCel™, and Eshmuno® HCX. 
     
     
         12 . A purified biomolecule obtained by the method according to any one of  claims 1  to  11 . 
     
     
         13 . A preparation comprising:
 (a) a biomolecule; and   (b) an aqueous buffer solution comprising one or more of:
 (i) at least 15 mM Good's buffer, 
 (ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine; 
 (iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and 
 (iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol. 
   
     
     
         14 . The preparation according to  claim 13 , wherein the aqueous buffer solution comprises 15 mM and 2.5 M Good's buffer. 
     
     
         15 . The preparation according to  claim 13  or  14 , wherein the at least one Good's buffer is selected from the group consisting of MES, MOPS, or Tris. 
     
     
         16 . The preparation according to  claim 13  or  14 , wherein the Good's buffer is MES. 
     
     
         17 . The preparation according to  claim 13  or  14 , wherein the Good's buffer is MOPS. 
     
     
         18 . The preparation according to any one of  claims 13  to  17 , wherein the biomolecule is a protein. 
     
     
         19 . The preparation according to  claim 18 , wherein the protein is an enzyme. 
     
     
         20 . The preparation according to  claim 19 , wherein the enzyme is a glycosidase. 
     
     
         21 . The preparation according to  claim 20 , wherein the glycosidase is an alpha-galactosidase. 
     
     
         22 . The preparation according to  claim 21 , wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta. 
     
     
         23 . A method for desorbing a biomolecule from a multimodal resin, comprising contacting a biomolecule bound to a multimodal resin with an aqueous buffer, wherein said aqueous buffer solution comprises one or more of:
 (i) at least 15 mM Good's buffer;   (ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;   (iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and   (iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol.   
     
     
         24 . A method for increasing the recovery rate of a biomolecule adsorbed on a multimodal resin by contacting said multimodal resin onto which said biomolecule has been adsorbed with an aqueous buffer solution, wherein said aqueous buffer solution comprises one or more of:
 (i) at least 15 mM Good's buffer;   (ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;   (iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and   (iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol.   
     
     
         25 . The method of  claim 23  or  claim 24 , wherein the aqueous buffer solution comprises between 15 mM and 2.5 M Good's buffer. 
     
     
         26 . The method of any one of  claims 23  to  25 , wherein the Good's buffer is selected from the group consisting of MES, MOPS, and Tris. 
     
     
         27 . The method of any one of  claims 23  to  25 , wherein the Good's buffer is MES. 
     
     
         28 . The method of any one of  claims 23  to  25 , wherein the Good's buffer is MOPS. 
     
     
         29 . The method of any one of  claims 23  to  28 , wherein the biomolecule is a protein. 
     
     
         30 . The method of  claim 29 , wherein the protein is an enzyme. 
     
     
         31 . The method of  claim 30 , wherein the enzyme is a glycosidase. 
     
     
         32 . The method of  claim 31 , wherein the glycosidase is an alpha-galactosidase. 
     
     
         33 . The method of  claim 32 , wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta. 
     
     
         34 . The method of any one of  claims 23  to  33 , wherein the multimodal resin is selected from the group consisting of Capto™ Adhere, Capto™ MMC, Pall PPA HyperCel™, Pall HEA HyperCel™, Pall MEP HyperCel™, and Eshmuno® HCX. 
     
     
         35 . An article of manufacture comprising:
 (a) a packaging material or container:   (b) an aqueous buffer solution comprising one or more of:
 (i) at least 15 mM Good's buffer; 
 (ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine; 
 (iii) between 0 and 400 mM in total concentration of urea, guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and 
 (iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol; 
 and 
   (c) a data carrier containing instructions for carrying out a method according to any one of  claims 1  to  11 .   
     
     
         36 . The article of manufacture according to  claim 35 , wherein the data carrier is a label present on the packaging material or container or a packaging insert. 
     
     
         37 . The article of manufacture according to  claim 35  or  claim 36 , wherein the data carrier comprises written information, visual information, or computer-readable information. 
     
     
         38 . The article of manufacture according to any one of  claims 35  to  37 , wherein the data carrier further comprises one or more of the following information:
 (i) instructions for use of aqueous buffer solution; 
 (ii) quality information such as information about the lot/batch number, the manufacturing or assembly site or the expiry or sell-by date of the aqueous buffer solution; or information concerning the correct storage or handling of the article; 
 (iii) information concerning the composition of the aqueous buffer solution; 
 (iv) information concerning the interpretation of information obtained when performing methods according to any one of  claims 1  to  11 ; 
 (v) a warning concerning possible misinterpretations or wrong results when applying unsuitable methods; and 
 (vi) a warning concerning possible misinterpretations or wrong results when using unsuitable reagent(s) and/or buffer(s). 
 
     
     
         39 . The article of manufacture according to any one of  claims 35  to  38 , further comprising one or more of the following components:
 (d) a chromatography medium; 
 (e) a chromatography column; and 
 (f) a reference biomolecule usable for calibrating a method for purifying or enriching a biomolecule. 
 
     
     
         40 . The article of manufacture according to  claim 39 , wherein said chromatography column is prepacked with said chromatography medium. 
     
     
         41 . The article of manufacture according to any one of  claims 39  to  40 , wherein said chromatography medium is a multimodal chromatography resin. 
     
     
         42 . The article of manufacture according to  claim 41 , wherein the multimodal chromatography resin is selected from the group consisting of Capto™ Adhere, Capto™ MMC, Pall PPA HyperCel™, Pall HEA HyperCel™, Pall MEP HyperCel™, and Eshmuno® HCX. 
     
     
         43 . The article of manufacture according to any one of  claims 39  to  42 , wherein the reference biomolecule is a protein or a nucleic acid. 
     
     
         44 . The article of manufacture according to any one of  claims 35  to  43 , wherein the aqueous buffer solution comprises between 15 mM and 2.5 M Good's buffer. 
     
     
         45 . The article of manufacture according to any one of  claims 35  to  44 , wherein the at least one Good's buffer is selected from the group consisting of MES, MOPS, and Tris. 
     
     
         46 . The article of manufacture according to any one of  claims 35  to  45 , wherein the Good's buffer is MES. 
     
     
         47 . The article of manufacture according to any one of  claims 35  to  45 , wherein the Good's buffer is MOPS.