US2011313147A1PendingUtilityA1

Chromatographic material for the absorption of proteins at physiological ionic strength

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Assignee: BOSCHETTI EGISTOPriority: Jan 20, 2004Filed: Aug 16, 2011Published: Dec 22, 2011
Est. expiryJan 20, 2024(expired)· nominal 20-yr term from priority
Inventors:Egisto BoschettiPierre Girot
G01N 30/02G01N 2035/00158B01J 41/20B01J 2219/00637B01J 2219/00527Y10S435/803B01J 39/26B01J 2219/00626B01J 2219/00605B01J 2219/0061C07K 1/16G01N 33/54353B01D 15/361B01J 2219/00612B01J 20/3242B01J 2220/54B01D 15/362B01J 2219/0074B01J 20/289B01D 15/327C07K 1/18B01J 2219/00702B01J 20/3285
52
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Claims

Abstract

Ion exchange and hydrophobic interaction chromatographic materials are constructed by tethering a terminal binding functionality to a solid support via a hydrophobic linker. The backbone of the linker typically comprises sulfur-containing moieties. Suitable terminal binding functionalities are tertiary amines, quaternary ammonium salts, or hydrophobic groups. These chromatographic materials possess both hydrophobic and ionic character under the conditions prescribed for their use. The separation of proteins from crude mixtures at physiological ionic strength can be accomplished with a chromatographic material of this type by applying pH or ionic strength gradients, thereby effecting protein adsorption and desorption.

Claims

exact text as granted — not AI-modified
1 . A chromatographic material comprising:
 (a) a terminal binding functionality;   (b) a hydrophobic linker comprising at least one functionality that is different from the terminal binding functionality; and   (c) a solid support,   
       wherein
 the hydrophobic linker links the terminal binding functionality to the solid support;
 and 
 
 the chromatographic material is capable of binding bovine albumin at physiological ionic strength, wherein the chromatographic material has the following general formula I: 
 
       
         
           
           
               
               
           
         
       
       wherein
 R 1 , R 2 , R 4  and R 5 , at each occurrence, are independently selected from the group consisting of H, C 1-6 -alkyl, C 1-6 -alkoxy, C 1-6 -alkyl-C 1-6 -alkoxy, aryl, C 1-6 -alkaryl, —NR′C(O)R″, —C(O)NR′R″, and hydroxy,
 wherein R′ and R″ are independently selected from C 1-6 -alkyl, and 
 wherein no more than one of R 1  and R 2  is hydroxy, 
 
 R 6  is selected from the group consisting of H, C 1-6 -alkyl, aryl, C 1-6 -alkaryl, —C(O)OH, —S(O) 2 OH, and —P(O)(OH) 2 ; 
 R 3  and R 3′ , together with X and Y, respectively, may independently be absent or present, and if present, then R 3  and R 3′  are independently selected from the group consisting of H, C 1-6 -alkyl, C 1-6 -alkoxy, C 1-6 -alkyl-C 1-6 -alkoxy, aryl, and C 1-6 -alkaryl,
 wherein X and Y, independently of each other, represent anions; 
 
 het and het′ are heteroatom moieties independently selected from the group consisting of —O—, —S—, —S(O)—, and —S(O) 2 —; 
 a, a′, a″, and a′″ are independently selected from the integers 0 through 6; 
 b and b′ are independently 0 or 1; 
 c is 0 or 1, and if c is 1, then (R 3 )X is absent; 
 d and d′ are independently 0 or 1; and 
 the wavy line represents the solid support. 
 
     
     
         2 . (canceled) 
     
     
         3 . The chromatographic material according to  claim 1 , wherein at least one of a, a′, a″, and a′″ is 2. 
     
     
         4 . The chromatographic material according to  claim 3 , wherein at least two of a, a′, a″, and a′″ are 2. 
     
     
         5 . The chromatographic material according to  claim 4 , further wherein at least one of a, a′, a″, and a′″ is 3. 
     
     
         6 . The chromatographic material according to  claim 1 , wherein at least one of a, a′, a″, and a′″ is 3. 
     
     
         7 . The chromatographic material according to  claim 6 , wherein at least two of a, a′, a″, and a′″ are 3. 
     
     
         8 . The chromatographic material according to  claim 6 , wherein a is 3. 
     
     
         9 . The chromatographic material according to  claim 8 , wherein het is S and b is 1. 
     
     
         10 . The chromatographic material according to  claim 9 , wherein a′ is selected from the group consisting of 2, 3, 4, 5, and 6. 
     
     
         11 . The chromatographic material according to  claim 10 , wherein b′ is 0. 
     
     
         12 . The chromatographic material according to  claim 11 , wherein c and d are both 0. 
     
     
         13 . The chromatographic material according to  claim 12 , wherein d′ is 1 and (R 3′ )Y is absent. 
     
     
         14 . The chromatographic material according to  claim 12 , wherein d′ is 1 and (R 3′ )Y is present. 
     
     
         15 . The chromatographic material according to  claim 12 , wherein d′ is 0. 
     
     
         16 . The chromatographic material according to  claim 11 , wherein c is 1 and d is 1. 
     
     
         17 . The chromatographic material according to  claim 16 , wherein d′ is 1 and (R 3′ )Y is absent. 
     
     
         18 . The chromatographic material according to  claim 16 , wherein d′ is 1 and (R 3′ )Y is present. 
     
     
         19 . The chromatographic material according to  claim 16 , wherein d′ is 0. 
     
     
         20 . The chromatographic material according to  claim 10 , wherein R 1  and R 2  are independently selected from H and C 1-6 -alkyl. 
     
     
         21 . The chromatographic material according to  claim 20 , wherein each of R 1  and R 2  are H. 
     
     
         22 . The chromatographic material according to  claim 8 , wherein d′ is 1. 
     
     
         23 . The chromatographic material according to  claim 22 , wherein R 3′ , R 5 , and R 6  are independently selected from the group consisting of H, C 1-6 -alkyl, aryl, and C 1-6 -alkaryl. 
     
     
         24 . The chromatographic material according to  claim 23 , wherein R 3′ , R 5 , and R 6  are independently selected from C 1-6 -alkyl and aryl. 
     
     
         25 . The chromatographic material according to  claim 24 , wherein R 3′ , R 5 , and R 6  are independently selected from C 1-6 -alkyl. 
     
     
         26 . The chromatographic material according to  claim 25 , wherein R 3′ , R 5 , and R 6  are independently selected from methyl and ethyl. 
     
     
         27 . The chromatographic material according to  claim 24 , wherein one of a″ and a′″ is 1 and the other is 1 or 2. 
     
     
         28 . The chromatographic material according to  claim 25 , wherein (R 3′ )Y is absent. 
     
     
         29 . The chromatographic material according to  claim 8 , wherein d′ is 0. 
     
     
         30 . The chromatographic material according to  claim 29 , wherein R 6  is H, C 1-6 -alkyl, aryl, or C 1-6 -alkaryl. 
     
     
         31 . The chromatographic material according to  claim 30 , wherein R 6  is selected from C 1-6 -alkyl and aryl. 
     
     
         32 . The chromatographic material according to  claim 31 , wherein R 6  is phenyl. 
     
     
         33 . The chromatographic material according to  claim 31 , wherein one of a″ and a′″ is 1 and the other is 1 or 2. 
     
     
         34 . The chromatographic material according to  claim 29 , wherein R 6  is —C(O)OH, —S(O) 2 OH, and —P(O)(OH) 2 . 
     
     
         35 . The chromatographic material according to  claim 34 , wherein one of a″ and a′″ is 1 and the other is 1 or 2. 
     
     
         36 . The chromatographic material according to  claim 1 , wherein:
 a is 3;   a′ is 2;   b is 1; and   each R 1  and R 2  in (CR 1 R 2 ) a  and (CR 1 R 2 ) a′  is H, except that one of R 2  in (CR 1 R 2 ) a  is optionally OH.   
     
     
         37 . The chromatographic material according to  claim 36 , wherein one of R 2  in (CR 1 R 2 ) a  is OH. 
     
     
         38 . The chromatographic material according to  claim 37 , wherein each of a″, a′″, and b′ is 0. 
     
     
         39 . The chromatographic material according to  claim 1 , wherein:
 a″ is 3;   a′″ is 2;   b′ is 1; and   each R 1  and R 2  in (CR 1 R 2 ) a″  and (CR 1 R 2 ) a′″  is H, except that one of R 2  in (CR 1 R 2 ) a″  is optionally OH.   
     
     
         40 . The chromatographic material according to  claim 39 , wherein one of R 2  in (CR 1 R 2 ) a″  is OH. 
     
     
         41 . The chromatographic material according to  claim 40 , wherein each of a, a′, and b″ is 0. 
     
     
         42 . The chromatographic material according to  claim 1 , wherein:
 a is 3;   a′ is 3;   b is 1; and   each R 1  and R 2  in (CR 1 R 2 ) a  and (CR 1 R 2 ) a′  is H.   
     
     
         43 . The chromatographic material according to  claim 42 , wherein each of a″, a′″, and b′ is 0. 
     
     
         44 . The chromatographic material according to  claim 1 , wherein:
 a″ is 3;   a′″ is 3;   b′ is 1; and   each R 1  and R 2  in (CR 1 R 2 ) a″  and (CR 1 R 2 ) a′″  is H.   
     
     
         45 . The chromatographic material according to  claim 44 , wherein each of a, a′, and b is 0. 
     
     
         46 . The chromatographic material according to  claim 1 , wherein
 a is 3;   a′ is 5;   b is 1; and   each R 1  and R 2  in (CR 1 R 2 ) a  and (CR 1 R 2 ) a′  is H.   
     
     
         47 . The chromatographic material according to  claim 46 , wherein:
 b′ is 0;   one of a″ and a′″ is 2 or 3, the other being 0; and   each R 1  and R 2  in (CR 1 R 2 ) a″  and (CR 1 R 2 ) a′″  is H, except that one of R 2  in (CR 1 R 2 ) a″  and (CR 1 R 2 ) a′″  is optionally OH.   
     
     
         48 . The chromatographic material according to  claim 47 , wherein a″ or a′″ is 3 and one of R 2  in (CR 1 R 2 ) a″  and (CR 1 R 2 ) a′″  is OH. 
     
     
         49 . The chromatographic material according to  claim 1 , wherein a or a′ is 3, the other being 0;
 a″ or a′″ is 3; 
 each of b, b′, and c is 0; and 
 d is 1. 
 
     
     
         50 . The chromatographic material according to  claim 49 , wherein d′ is 1. 
     
     
         51 . The chromatographic material according to  claim 1 , wherein:
 each of R 1  and R 2  are H;   R 3 , R 3′ , R 4 , R 5 , and R 6  are independently selected from the group consisting of H, C 1-6 -alkyl, aryl, and C 1-6 -alkaryl;   het is S;   a is 3;   a′ is selected from the group consisting of 2, 3, 4, 5, and 6;   one of a″ and a′″ is 1 and the other is 1 or 2; and   b is 1 and b′ is 0.   
     
     
         52 . The chromatographic material according to  claim 1 , selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         53 . The chromatographic material according to  claim 1 , wherein the wherein the solid support is an organic material. 
     
     
         54 . The chromatographic material according to  claim 53 , wherein the organic material is one selected from the group consisting of cellulose, agarose, dextran, polyacrylates, polystyrene, polyacrylamide, polymethacrylamide, copolymers of styrene and divinylbenzene, and mixtures thereof. 
     
     
         55 . The chromatographic material according to  claim 1 , wherein the solid support is an inorganic material. 
     
     
         56 . The chromatographic material according to  claim 55 , wherein the inorganic material is one selected from the group consisting of silica, zirconia, alumina, titania, ceramics, and mixtures thereof. 
     
     
         57 . The chromatographic material according to  claim 1 , wherein the solid support is in the form of a bead or particle. 
     
     
         58 . The chromatographic material according to  claim 1 , wherein the solid support is a planar solid support. 
     
     
         59 - 98 . (canceled) 
     
     
         99 . The chromatographic material of  claim 1 , wherein
 R 1 , R 2 , R 4 , and R 5 , at each occurrence, are independently selected from the group consisting of H, C 1-6 -alkyl, C 1-6 -alkoxy, C 1-6 -alkyl-C 1-6 -alkoxy, —NR′C(O)R″, —C(O)NR′R″, and hydroxy,
 wherein R′ and R″ are independently selected from C 1-6 -alkyl, and 
 wherein no more than one of R 1  and R 2  is hydroxy; and, 
   R 6  is selected from the group consisting of H, C 1-6 -alkyl, —C(O)OH, —S(O) 2 OH, and —P(O)(OH) 2 .   
     
     
         100 . The chromatographic material of  claim 1 , wherein
 R 6  is selected from the group consisting of C 1-6 -alkyl, —C(O)OH, —S(O) 2 OH, and —P(O)(OH) 2 .   
     
     
         101 . The chromatographic material of  claim 1 , wherein
 R 1 , R 2 , R 4 , and R 5 , at each occurrence, are independently selected from the group consisting of H, C 1-6 -alkyl, C 1-6 -alkoxy, C 1-6 -alkyl-C 1-6 -alkoxy, aryl, C 1-6 -alkaryl, and hydroxy,   wherein no more than one of R 1  and R 2  is hydroxy;   R 6  is selected from the group consisting of H, C 1-6 -alkyl, aryl, and C 1-6 -alkaryl;   R 3  and R 3′ , together with X and Y, are both absent; and   d and d′ are both 0.   
     
     
         102 . The chromatographic material of  claim 1 , wherein
 a is 3;   a″ is 0 or 3;   d is 1;   a′, a′″, b, b′, c, and d′ are all 0;   (R 3 )X is absent;   each R 1 , R 2 , and R 4 , is H; and   R 6  is selected from the group consisting of C 1-6 -alkyl and aryl.   
     
     
         103 . The chromatographic material of  claim 1 , wherein
 a is 3;   d is 1;   a′, a″, a′″, b, b′, c, and d′ are all 0;   (R 3 )X is absent;   each R 1 , R 2 , and R 4 , is H; and   R 6  is selected from the group consisting of C 1-6 -alkyl.   
     
     
         104 . The chromatographic material of  claim 1 , wherein
 a is 3;   a″ is 3;   d is 1;   a′, a′″, b, b′, c, and d′ are all 0;   (R 3 )X is absent;   R 1  and R 2  at each occurrence, are independently selected from the group consisting of H and hydroxy;   wherein no more than one of R 1  and R 2  is hydroxy;   R 4  is H; and   R 6  is selected from the group consisting of aryl.   
     
     
         105 . The chromatographic material of  claim 104 , wherein
 a is 3;   a″ is 3;   d is 1;   a′, a′″, b, b′, c, and d′ are all 0;   (R 3 )X is absent;   each R 1 , R 2 , and R 4 , is H; and   R 6  is selected from the group consisting of aryl.   
     
     
         106 . The chromatographic material of  claim 104 , wherein R 6  is phenyl. 
     
     
         107 . The chromatographic material of  claim 105 , wherein R 6  is phenyl.

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