US2020406229A1PendingUtilityA1

Analytical method for sugar chains having acidic groups

45
Assignee: JAPAN CHEM RESPriority: Nov 15, 2017Filed: Nov 14, 2018Published: Dec 31, 2020
Est. expiryNov 15, 2037(~11.3 yrs left)· nominal 20-yr term from priority
Inventors:Tomoki FukatsuChika Morooka
G01N 30/461G01N 30/34C07K 16/00B01J 41/20B01J 41/14B01J 41/07B01J 41/05B01J 20/3248B01J 20/3219B01J 20/286B01D 15/363B01D 15/305B01D 15/1871G01N 2030/8836G01N 2030/067G01N 30/46G01N 30/74G01N 30/88G01N 30/06B01J 20/283B01J 20/285
45
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

A chromatography column for the use of separation of acidic sugar chains, wherein the column comprises a first column and a second column, the second column connected by a flow path downstream of an outlet of the first column, and selected from the following (1) or (2): (1) the carrier of the first column is hydrophobically modified silica having a group containing a primary amine, a secondary amine or/and a tertiary amine, and the carrier of the second column is a resin having a group containing a primary amine, a secondary amine or/and a tertiary amine; (2) the carrier of the first column is a resin having a group containing a primary amine, a secondary amine or/and a tertiary amine, and the carrier of the second column is hydrophobically modified silica having a group containing a primary amine, a secondary amine, or/and a tertiary amine.

Claims

exact text as granted — not AI-modified
1 . A chromatography column comprising:
 a first column having an outlet; and   a second column connected downstream of the outlet of the first column,   wherein the first column and the second column satisfy either (1) or (2):   (1) the first column includes a carrier 1 which is hydrophobically modified silica having a group including at least one of a primary amine, a secondary amine and a tertiary amine, and the second column includes a carrier 2 which is a resin having a group including at least one of a primary amine, a secondary amine and a tertiary amine; or   (2) the first column includes the carrier 2, and the second column includes the carrier 1.   
     
     
         2 . The chromatography column according to  claim 1 , wherein the first column and the second column are hydrophilic interaction chromatography columns. 
     
     
         3 . The chromatography column according to  claim 1 , wherein the carrier 2 is an anion exchange resin. 
     
     
         4 . The chromatography column according to  claim 1 , wherein the carrier 1 has a hydrophobic group of formula I and an amino group of formula II: 
       
         
           
           
               
               
           
         
       
       
         
           
           
               
               
           
         
         where in the formula I, X represents a silica portion of the carrier 1, and 
         in the formula II, X represents the silica portion of the carrier 1, and R represents an alkyl of —(CH 2 )n— where n=1-8. 
       
     
     
         5 . The chromatography column according to  claim 1 , wherein the carrier 2 has an amino group of formula III,
 formula III
   Y—NH(CH 2 CH 2 NH) n H
 
   where Y represents a resin portion of the carrier 2, and n is an integer of 1 to 8.   
     
     
         6 . The chromatography column according to  claim 1 , wherein the resin is made from polyvinyl alcohol. 
     
     
         7 . A method for separating an acidic sugar chain from a sample, comprising:
 labeling the acidic sugar chain in the sample with an aminopyridine;   subjecting the sample including an aminopyridine-labeled acidic sugar chain to chromatography on the chromatography column of  claim 1 ;   continuously directing an effluent from the chromatography column to a flow path;   continuously measuring fluorescence intensity of the effluent flowing in the flow path;   and   identifying a peak of fluorescence intensity corresponding to the acidic sugar chain in a chromatogram obtained,   wherein the subjecting of the sample to the chromatography includes passing a first mobile phase through the chromatography column and thereafter passing a second mobile phase through the chromatography column, and the second mobile phase has hydrophobicity lower than hydrophobicity of the first mobile phase.   
     
     
         8 . The method according to  claim 7 , wherein the acidic sugar chain is from a glycoprotein in the sample. 
     
     
         9 . The method according to  claim 8 , wherein the glycoprotein is a lysosomal enzyme. 
     
     
         10 . The chromatography column according to  claim 2 , wherein the carrier 2 is an anion exchange resin. 
     
     
         11 . The chromatography column of according to  claim 2 , wherein the carrier 1 has a hydrophobic group of formula I and an amino group of formula II: 
       
         
           
           
               
               
           
         
         where in the formula I, X represents a silica portion of the carrier 1, and 
         in the formula II, X represents the silica portion of the carrier 1, and R represents an alkyl of —(CH 2 )n— where n=1-8. 
       
     
     
         12 . The chromatography column according to  claim 11 , wherein the carrier 2 has an amino group of formula III,
 formula III
   Y—NH(CH 2 CH 2 NH) n H
 
   where Y represents a resin portion of the carrier 2, and n is an integer of 1 to 8.   
     
     
         13 . The chromatography column according to  claim 4 , wherein the carrier 2 has an amino group of formula III,
 formula III
   Y—NH(CH 2 CH 2 NH) n H
 
   where Y represents a resin portion of the carrier 2, and n is an integer of 1 to 8.   
     
     
         14 . The chromatography column according to  claim 2 , wherein the resin is made from polyvinyl alcohol. 
     
     
         15 . A method for separating an acidic sugar chain from a sample, comprising:
 labeling the acidic sugar chain in the sample with an aminopyridine;   subjecting the sample including an aminopyridine-labeled acidic sugar chain to chromatography on the chromatography column of  claim 2 ;   continuously directing an effluent from the chromatography column to a flow path;   continuously measuring fluorescence intensity of the effluent flowing in the flow path; and   identifying a peak of fluorescence intensity corresponding to the acidic sugar chain in a chromatogram obtained,   wherein the subjecting of the sample to the chromatography includes passing a first mobile phase through the chromatography column and thereafter passing a second mobile phase through the chromatography column, and the second mobile phase has hydrophobicity lower than hydrophobicity of the first mobile phase.   
     
     
         16 . The method according to  claim 15 , wherein the acidic sugar chain is from a glycoprotein in the sample. 
     
     
         17 . The method according to  claim 16 , wherein the glycoprotein is a lysosomal enzyme. 
     
     
         18 . A method for separating an acidic sugar chain from a sample, comprising:
 labeling the acidic sugar chain in the sample with an aminopyridine;   subjecting the sample including an aminopyridine-labeled acidic sugar chain to chromatography on the chromatography column of  claim 12 ;   continuously directing an effluent from the chromatography column to a flow path;   continuously measuring fluorescence intensity of the effluent flowing in the flow path; and   identifying a peak of fluorescence intensity corresponding to the acidic sugar chain in a chromatogram obtained,   wherein the subjecting of the sample to the chromatography includes passing a first mobile phase through the chromatography column and thereafter passing a second mobile phase through the chromatography column, and the second mobile phase has hydrophobicity lower than hydrophobicity of the first mobile phase.   
     
     
         19 . The method according to  claim 18 , wherein the acidic sugar chain is from a glycoprotein in the sample. 
     
     
         20 . The method according to  claim 19 , wherein the glycoprotein is a lysosomal enzyme.

Join the waitlist — get patent alerts

Track US2020406229A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.