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SUGAR-CHAIN CONCENTRATION COLUMN AND SUGAR-CHAIN CONCENTRATION METHOD

Foreign code F180009311
File No. (S2016-0753-N0)
Posted date Jan 24, 2018
Country WIPO
International application number 2017JP018037
International publication number WO 2017195887
Date of international filing May 12, 2017
Date of international publication Nov 16, 2017
Priority data
  • P2016-095997 (May 12, 2016) JP
  • P2016-169200 (Aug 31, 2016) JP
Title SUGAR-CHAIN CONCENTRATION COLUMN AND SUGAR-CHAIN CONCENTRATION METHOD
Abstract Disclosed is a novel sugar-chain concentration means that is capable of handling a minute amount of a sample and increasing the concentration of a glycopeptide or a free sugar chain within a short period of time. This sugar-chain concentration column includes, in the column, a solid-phase extraction carrier layer, and a polymeric polyol layer directly or indirectly layered on the solid-phase extraction carrier layer. An octadecyl silyl silica gel or the like, and cellulose fine crystals or the like can be suitably used as the solid-phase extraction carrier and the polymeric polyol, respectively. The column can be used for increasing all concentrations of N-bound type sugar chains, O-bound type sugar chains, and free sugar chains.
Scope of claims [claim1]
1. In each column, and a solid phase extraction by a carrier layer, directly or indirectly on the stacked layer including a polymeric polyols, sugar chain concentrated column.
[claim2]
2. Wherein the carrier is a solid-phase extraction as at least one inverse phase resin 1, according to claim 1 column.
[claim3]
3. Wherein the carrier is a solid-phase extraction, octadecylsilyl-silica gel, porous solid carbon material, as well as the monomers, divinylbenzene, N containing methacrylate, and 1 is selected from one of the at least N- vinyl pyrrolidone polymer having functional groups selected from one of at least 1, claims 1 or 2 column.
[claim4]
4. Wherein the carrier is a solid-phase extraction, octadecylsilyl-silica gel, porous solid carbon material, styrene divinylbenzene polymer, divinyl benzene polymer, a methacrylate containing N, and poly (N - vinylpyrrolidon) at least one selected from 1, according to claim 3 column.
[claim5]
5. Wherein the polymer is a polysaccharide polymer polyol, of any one of claims 1-4 column.
[claim6]
6. Polysaccharide polymers, cellulose, agarose, dextran and at least one selected from 1, according to claim 5 column.
[claim7]
7. Microcrystalline cellulose cellulose or cellulose fiber, according to claim 6 column.
[claim8]
8. A carrier layer with solid-phase extraction and polymeric polyols in a volume ratio of 1:12.5-160, of any one of claims 1-7 column.
[claim9]
9. Column capacity in 100 μl-500 μl, of any one of claims 1-8 column.
[claim10]
10. The volume of the layer of polymeric polyols in 5-50 μl, of any one of claims 1-9 column.
[claim11]
11. Any one of claims 1-10 concentrated sugar chain of column dissolved sugar chain containing samples and the sample, and dilution, polymeric polyol layer from the solid-phase extraction column in a direction toward the carrier layer is applied to the pressure, glycopeptide or free sugar chain is stored in the layer of polymeric polyols, a sample-adding step; cleaning liquid to the column and the column, polymeric polyol layer from the solid-phase extraction column in a direction toward the carrier layer is applied to the pressure of the column by washing, to remove the contaminants within the layer of polymeric polyols Concentrate sugar chain peptides or free sugar is held, the step of concentrating selectively; and eluent to the column and, polymeric polyol layer by solid phase extraction from the carrier layer in a direction of the pressure applied to the column elution of a peptide, comprising the steps of elution, glycopeptide or free of the sugar chain method concentrated, dissolved sample, a diluting liquid, the cleaning liquid column, in which a polar organic solvent and water as the eluent, method.
[claim12]
12. A glycopeptide method concentrated, in the step of concentrating then selectively, to the column and aqueous cleaning, polymeric polyol layer from the solid-phase extraction column in a direction toward the carrier layer is applied to the pressure, glycopeptide solid-phase extraction mixtureto carrier layer, the step of concentrating re is performed, the method according to claim 11.
[claim13]
13. In which a polar organic solvent is water, the concentration of the polar organic solvent is 75% -95%, the method according to claim 12.
[claim14]
14. A method of isolated sugar chain concentrated, dissolved sample, dilution and polar organic solvent is a water column used as a cleaning liquid, the concentration of 75% -95% and a polar organic solvent, a polar organic solvent is used as the eluent water, the concentration of the polar organic solvent is 40% -60%, the method according to claim 11.
[claim15]
15. A polar organic solvent, and a lower alcohol is selected from acetonitrile and at least one of 1, the method of any one of claims 11-14.
[claim16]
16. The application of pressure to the column is carried out by centrifugation, method according to any one of claims 11-15.
[claim17]
17. Including a column of any one of claims 1-10, sugar chain concentrated kit.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • YOKOHAMA CITY UNIVERSITY
  • Inventor
  • OHTA YUKI
  • KAWASAKI NANA
  • TAKAKURA DAISUKE
IPC(International Patent Classification)
Specified countries (WO2017195887)
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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