Method of separating biomolecules using hydrophobically-derivatized supports

Abstract

A method of separating biomolecules in an aqueous mixture is disclosed comprising a obtaining a separation vessel containing separation media, wherein the separation media comprises a porous support with a hydrophobic monomer grafted thereon, the hydrophobic monomer having the structure: CH 2 ═CR 4 C(O)NHC(R 1 R 1 )(C(R 1 R 1 )) n C(O)XR 3 wherein n is an integer of 0 or 1; R 1 is independently selected from at least one of: a hydrogen atom, alkyls, aryls, and alkylaryls, wherein the alkyls, aryls, and alkylaryls have a total of 10 carbon atoms or less; R 3 is a hydrophobic group selected from at least one of alkyls, aryls, alkylaryls and ethers, wherein the alkyls, aryls, alkylaryls and ethers have a total number of carbon atoms ranging from 4 to 30; R 4 is H or CH 3 ; and X is O or NH; wherein the hydrophobic monomer is derived from an amine or an alcohol (HXR 3 ) that has a hydrophilicity index of 25 or less; and (b) passing the aqueous mixture through the separation vessel thereby separating the biomolecules. Such methods can be used to separate proteins, antibodies, fusion proteins, vaccines, peptides, enzymes, DNA, and/or RNA.

We claim: 
     
         1 . A method of separating a biomolecule in an aqueous mixture, the method comprising:
 (a) obtaining a separation vessel containing separation media, wherein the separation media comprises a porous support with a hydrophobic monomer grafted thereon, the hydrophobic monomer having the structure:
   CH 2 ═CR 4 C(O)NHC(R 1 R 1 )(C(R 1 R 1 )) n C(O)XR 3  
 
   wherein n is an integer of 0 or 1; R 1  is independently selected from at least one of:   a hydrogen atom, alkyls, aryls, and alkylaryls, wherein the alkyls, aryls, and alkylaryls have a total of 10 carbon atoms or less; R 3  is a hydrophobic group selected from at least one of alkyls, aryls, alkylaryls and ethers, wherein the alkyls, aryls, alkylaryls and ethers have a total number of carbon atoms ranging from 4 to 30; R 4  is H or CH 3 ; and X is O or NH; wherein the hydrophobic monomer is derived from an amine or an alcohol (HXR 3 ) that has a hydrophilicity index of 25 or less; and   (b) passing the aqueous mixture through the separation vessel thereby separating the biomolecule.   
     
     
         2 . The method of  claim 1 , wherein R 1  is independently selected from at least one of: methyl, ethyl, phenyl, or combinations thereof. 
     
     
         3 . The method of  claim 1 , wherein R 3  is selected from at least one of: benzyl, phenethyl, phenoxyethyl, phenylpropyl, butyl, pentyl, hexyl, octyl, dodecyl, octadecyl, phenylbutyl, or combinations thereof. 
     
     
         4 . The method of  claim 1 , wherein R 3  is selected from: benzyl, phenethyl, phenoxyethyl, phenylpropyl, butyl, pentyl, hexyl, octyl, phenylbutyl, or combinations thereof. 
     
     
         5 . The method of  claim 1 , wherein n is 1. 
     
     
         6 . The method of  claim 1 , wherein the hydrophobic monomer has the following structure:
   CH 2 ═CR 4 C(O)NHC(R 2 R 2 )(C(R 1 R 1 )) n C(O)NHR 3  
   
       wherein n is an integer of 0 or 1; wherein each R 2  is independently selected from: alkyls, aryls, and alkylaryls, wherein the alkyls, aryls, and alkylaryls have a total of 10 carbon atoms or less; each R 1  is independently selected from: alkyls, aryls, and alkylaryls, wherein the alkyls, aryls, and alkylaryls have a total of 10 carbon atoms or less; R 3  is a hydrophobic group selected from: alkyls, aryls, alkylaryls and ethers, wherein the alkyls, aryls, alkylaryls and ethers have a total number of carbon atoms ranging from 4 to 30; and R 4  is H or CH 3 . 
     
     
         7 . The method of  claim 6 , wherein R 2  is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and ethylhexyl. 
     
     
         8 . The method of  claim 6 , wherein R 2  is selected from: methyl or ethyl. 
     
     
         9 . The method of  claim 6 , wherein R 2  is the same. 
     
     
         10 . The method of  claim 6 , wherein n is 1. 
     
     
         11 . The method of  claim 1 , wherein the hydrophobic monomer has a structure selected from at least one of:
 CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 4 C 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)O(CH 2 ) 4 C 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NHCH 2 C 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 2 C 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 2 OC 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 3 C 6 H 5 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 3 CH 3 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 5 CH 3 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 7 CH 3 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 11 CH 3 ;   CH 2 ═CHC(O)NHC(CH 3 )(CH 3 )C(O)NH(CH 2 ) 17 CH 3 ; or combinations thereof.   
     
     
         12 . The method of  claim 1 , wherein the porous support is selected from a woven web, a nonwoven web, a fibrous web, a microporous membrane, a microporous film, and combinations thereof. 
     
     
         13 . The method of  claim 1 , wherein the biomolecule is selected from at least one of: proteins, antibodies, fusion proteins, vaccines, peptides, enzymes, DNA, or RNA. 
     
     
         14 . The method of  claim 1 , wherein the biomolecule is a biomacromolecule. 
     
     
         15 . The method of  claim 1 , wherein the aqueous mixture comprises a buffer. 
     
     
         16 . The method of  claim 1 , wherein the aqueous mixture is a biological sample.