CN110130099B - Magnetic nanofiber-based zwitterionic hydrophilic materials for selective capture and recognition of glycopeptides - Google Patents
Magnetic nanofiber-based zwitterionic hydrophilic materials for selective capture and recognition of glycopeptides Download PDFInfo
- Publication number
- CN110130099B CN110130099B CN201910340139.3A CN201910340139A CN110130099B CN 110130099 B CN110130099 B CN 110130099B CN 201910340139 A CN201910340139 A CN 201910340139A CN 110130099 B CN110130099 B CN 110130099B
- Authority
- CN
- China
- Prior art keywords
- mhn
- nanofiber
- gsh
- nanoparticles
- glycopeptides
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 239000002121 nanofiber Substances 0.000 title claims abstract description 99
- 102000002068 Glycopeptides Human genes 0.000 title claims abstract description 72
- 108010015899 Glycopeptides Proteins 0.000 title claims abstract description 72
- 239000000463 material Substances 0.000 title claims abstract description 37
- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 claims abstract description 42
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims abstract description 28
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 26
- 239000002105 nanoparticle Substances 0.000 claims abstract description 25
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims abstract description 10
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims abstract description 10
- 230000003993 interaction Effects 0.000 claims abstract description 6
- RWSXRVCMGQZWBV-PHDIDXHHSA-N L-Glutathione Natural products OC(=O)[C@H](N)CCC(=O)N[C@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-PHDIDXHHSA-N 0.000 claims abstract description 5
- 230000004048 modification Effects 0.000 claims abstract description 5
- 238000012986 modification Methods 0.000 claims abstract description 5
- 125000003396 thiol group Chemical group [H]S* 0.000 claims abstract description 5
- 239000000203 mixture Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- XYYVDQWGDNRQDA-UHFFFAOYSA-K trichlorogold;trihydrate;hydrochloride Chemical compound O.O.O.Cl.Cl[Au](Cl)Cl XYYVDQWGDNRQDA-UHFFFAOYSA-K 0.000 claims description 4
- 229910001868 water Inorganic materials 0.000 claims description 4
- CDVAIHNNWWJFJW-UHFFFAOYSA-N 3,5-diethoxycarbonyl-1,4-dihydrocollidine Chemical compound CCOC(=O)C1=C(C)NC(C)=C(C(=O)OCC)C1C CDVAIHNNWWJFJW-UHFFFAOYSA-N 0.000 claims description 3
- 239000012279 sodium borohydride Substances 0.000 claims description 3
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- ZIBGPFATKBEMQZ-UHFFFAOYSA-N triethylene glycol Chemical compound OCCOCCOCCO ZIBGPFATKBEMQZ-UHFFFAOYSA-N 0.000 claims description 3
- 229940071240 tetrachloroaurate Drugs 0.000 claims description 2
- 150000004684 trihydrates Chemical class 0.000 claims description 2
- 229960003180 glutathione Drugs 0.000 abstract description 79
- 238000001514 detection method Methods 0.000 abstract description 11
- 230000035945 sensitivity Effects 0.000 abstract description 8
- 238000011084 recovery Methods 0.000 abstract description 5
- 239000010931 gold Substances 0.000 description 30
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 18
- 238000000034 method Methods 0.000 description 18
- 238000004458 analytical method Methods 0.000 description 17
- 238000001228 spectrum Methods 0.000 description 14
- 210000002966 serum Anatomy 0.000 description 11
- 108090001008 Avidin Proteins 0.000 description 9
- 241000287828 Gallus gallus Species 0.000 description 9
- 230000008569 process Effects 0.000 description 9
- 108060003951 Immunoglobulin Proteins 0.000 description 8
- 239000007853 buffer solution Substances 0.000 description 8
- 102000018358 immunoglobulin Human genes 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 108010024636 Glutathione Proteins 0.000 description 7
- 230000029087 digestion Effects 0.000 description 7
- 238000001819 mass spectrum Methods 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 6
- 108090000288 Glycoproteins Proteins 0.000 description 6
- 102000003886 Glycoproteins Human genes 0.000 description 6
- 238000003917 TEM image Methods 0.000 description 6
- 238000002441 X-ray diffraction Methods 0.000 description 6
- 229940098773 bovine serum albumin Drugs 0.000 description 6
- 238000002149 energy-dispersive X-ray emission spectroscopy Methods 0.000 description 6
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- 239000003480 eluent Substances 0.000 description 5
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 102000004142 Trypsin Human genes 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical group O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- 239000004201 L-cysteine Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 239000000090 biomarker Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000002013 hydrophilic interaction chromatography Methods 0.000 description 3
- 229920002521 macromolecule Polymers 0.000 description 3
- 230000005415 magnetization Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000031018 biological processes and functions Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000000151 deposition Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 239000002122 magnetic nanoparticle Substances 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 238000000816 matrix-assisted laser desorption--ionisation Methods 0.000 description 2
- 229920002189 poly(glycerol 1-O-monomethacrylate) polymer Polymers 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 230000004043 responsiveness Effects 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 239000013177 MIL-101 Substances 0.000 description 1
- 102000000447 Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Human genes 0.000 description 1
- 108010055817 Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Proteins 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 125000005620 boronic acid group Chemical class 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000010267 cellular communication Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229910052681 coesite Inorganic materials 0.000 description 1
- 229910052906 cristobalite Inorganic materials 0.000 description 1
- 230000022811 deglycosylation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000007306 functionalization reaction Methods 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 229910021389 graphene Inorganic materials 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000000155 isotopic effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 238000007885 magnetic separation Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001254 matrix assisted laser desorption--ionisation time-of-flight mass spectrum Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012621 metal-organic framework Substances 0.000 description 1
- 229910052961 molybdenite Inorganic materials 0.000 description 1
- 229910052982 molybdenum disulfide Inorganic materials 0.000 description 1
- 239000002070 nanowire Substances 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229920000314 poly p-methyl styrene Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000013309 porous organic framework Substances 0.000 description 1
- 238000000634 powder X-ray diffraction Methods 0.000 description 1
- 238000004094 preconcentration Methods 0.000 description 1
- 206010063401 primary progressive multiple sclerosis Diseases 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 238000004729 solvothermal method Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 229910052682 stishovite Inorganic materials 0.000 description 1
- 229910052905 tridymite Inorganic materials 0.000 description 1
Images
Classifications
-
- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M11/00—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising
- D06M11/32—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with oxygen, ozone, ozonides, oxides, hydroxides or percompounds; Salts derived from anions with an amphoteric element-oxygen bond
- D06M11/36—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with oxygen, ozone, ozonides, oxides, hydroxides or percompounds; Salts derived from anions with an amphoteric element-oxygen bond with oxides, hydroxides or mixed oxides; with salts derived from anions with an amphoteric element-oxygen bond
- D06M11/49—Oxides or hydroxides of elements of Groups 8, 9,10 or 18 of the Periodic Table; Ferrates; Cobaltates; Nickelates; Ruthenates; Osmates; Rhodates; Iridates; Palladates; Platinates
-
- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M11/00—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising
- D06M11/83—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with metals; with metal-generating compounds, e.g. metal carbonyls; Reduction of metal compounds on textiles
-
- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M13/00—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with non-macromolecular organic compounds; Such treatment combined with mechanical treatment
- D06M13/322—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with non-macromolecular organic compounds; Such treatment combined with mechanical treatment with compounds containing nitrogen
- D06M13/402—Amides imides, sulfamic acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4044—Concentrating samples by chemical techniques; Digestion; Chemical decomposition
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
- G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Textile Engineering (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Computational Biology (AREA)
- General Physics & Mathematics (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Optics & Photonics (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a magnetic nanofiber-based zwitter-ion hydrophilic material for selectively capturing and identifying glycopeptides, which is used for enriching and identifying the glycopeptides; one-dimensional Hydroxyapatite Nanofiber (HN) is used as a carrier to fix Fe3O4Nanoparticles and Au nanoparticles, via the thiol group of the zwitterionic tripeptide L-glutathione with Au and Fe3O4The surface modification is carried out through the affinity interaction between the two to form mHN/Au-GSH nano-fiber. mHN/Au-GSH nanofiber has high glycopeptide detection sensitivity (2fmol), high recovery rate (89.65%), strong binding capacity (100mg/g) and high enrichment selectivity (1: 100).
Description
Technical Field
The invention relates to a magnetic nanofiber-based zwitter-ion hydrophilic material for selectively capturing and identifying glycopeptides, and belongs to the technical field of biochemical analysis.
Background
Proteins/peptides, carbohydrates, nucleic acids, etc. as essential and important biological components mediate most of the fundamental biological processes such as cellular communication, immune reactions and enzymatic reactions. Due to the complex composition and severe interference, the detection and characterization of target biological macromolecules is a great challenge, especially for low abundance analytes. Thus, sample preparation is an indispensable step in the analytical process. The development of affinity materials for selective separation and preconcentration of target biological macromolecules is an effective method to reduce impurity interference and improve detection sensitivity. To date, a variety of materials have been developed, including silica, mesoporous silica, magnetic nanoparticles, Au nanoparticles, polymers, graphene oxide, metal organic frameworks, and porous organic frameworks, for biomacromolecule extraction. The design and synthesis of affinity materials for highly selective capture and identification of target biological macromolecules remains an important research topic.
Glycoproteins are involved in a variety of biological processes and are also biomarkers for several diseases. The comprehensive identification of the glycoprotein not only can understand the physiological process, but also is beneficial to the discovery of biomarkers. The current popular glycopeptide omics analysis method based on mass spectrum comprises proteolysis, glycopeptide enrichment and mass spectrum identification. Based on the structural differences between glycopeptides and non-glycopeptides, several affinity materials including lectins, hydrazine beads, boronic acids, and hydrophilic adsorbents have been introduced to identify and separate glycopeptides and non-glycopeptides abundant in living organisms. In contrast, the hydrophilic interaction separation method has the advantages of high enrichment efficiency, wide glycan specificity, irreversible change of glycan components and the like.
Many hydrophilic materials have been proposed for the enrichment and identification of glycopeptides. Notably, various hydrophilic materials have been combined with magnetic nanoparticles to achieve magnetic response capabilities, making the separation process simple and efficient.
However, the synthesis of these hydrophilic materials often requires complex synthetic procedures, multi-step functionalization, and longer incubation times. This cumbersome process greatly increases the cost of the analytical methods, greatly limiting their application to practical samples. Therefore, more hydrophilic materials with abundant structural features need to be excavated to realize simple and efficient enrichment of glycopeptides.
Disclosure of Invention
The invention aims at the problems, thereby providing a magnetic nanofiber-based zwitter-ion hydrophilic material for selectively capturing and identifying glycopeptides, which has the characteristics of good hydrophilicity, abundant zwitter-ion glutathione molecules and strong magnetic responsiveness, and has better specificity and efficiency in the aspect of glycopeptide enrichment.
The specific technical scheme is as follows:
magnetic nanofiber-based zwitterion hydrophilic material for selectively capturing and identifying glycopeptides, and hydroxyapatite nanofiber is used as a carrier to fix Fe3O4Nanoparticles and Au nanoparticles, via the thiol group of the zwitterionic tripeptide L-glutathione with Au and Fe3O4The surface modification is carried out through the affinity interaction between the two to form mHN/Au-GSH nano-fiber.
Further, the preparation method of the mHN/Au-GSH nano fiber comprises the following steps:
s1: dispersing 2.0g of hydroxyapatite nanofiber and 0.75g of ferric acetylacetonate in 400mL of triethylene glycol to obtain a mixture;
s2: the mixture was transferred to a 1L autoclave and reacted at 220 ℃ for 12 hours, washed with water and ethanol to give fixed Fe3O4A nanofiber product of nanoparticles mHN;
s3: 0.5g of prepared mHN and 25mL of tetrachloroaurate trihydrate were added to 1L of deionized water to obtain a suspension;
s4: the suspension was stirred at 25 ℃ for 12 hours, 50mg of sodium borohydride was added and stirred for 30 minutes to obtain simultaneous fixed Fe3O4Nanofiber product mHN/Au of nanoparticles and Au nanoparticles;
s5: after washing with deionized water, 0.5g of the HN/Au product was dispersed in an aqueous solution containing 2g of glutathione; stirring at 50 deg.C for 10 hr to obtain mHN/Au-GSH nanofiber.
Further, the concentration of tetrachloroauric acid trihydrate in S3 was 2 mg/mL.
Further, mHN/Au-GSH nanofiber is used for enrichment and identification of glycopeptide.
Furthermore, the mHN/Au-GSH nano-fiber can be used for pretreatment before ionization time-of-flight mass spectrometry analysis of human immunoglobulin, chicken avidin protein or bovine serum albumin; the method comprises the following specific steps:
s1: digesting human immunoglobulin, chicken avidin or bovine serum albumin with trypsin;
s2: 400 u g mHN/Au-GSH nanofibers and 2.5 u g human immunoglobulin digests, chicken avidin digests or bovine serum albumin digests into 400 u L buffer solution, at 25 degrees C temperature in 15 minutes;
s3: magnetically separating the material, washing with 2 × 400 μ L buffer solution, and adding 20 μ L eluent to elute and capture glycopeptide;
s4: depositing 1 μ L of the eluate and 1 μ L of DHB on a MALDI plate, and performing MALDI-TOFMS analysis;
in the above pretreatment process, the buffer solution is ACN-H2O-TFA, 92: 7.9: 0.1, v/v/v; the eluent is ACN-H2O, 30: 70, v/v; the concentration of DHB was 25 mg/mL.
Further, the mHN/Au-GSH nanofiber can be used for pretreatment before nano-LC-MS/MS analysis of human serum; the method comprises the following specific steps:
s1: trypsinizing human serum;
s2: 2mg of mHN/Au-GSH nanofibers were incubated with 5. mu.L of human serum digest in 1mL of buffer for 30 min;
s3: adding 2 × 100 μ L of the eluate, and lyophilizing the obtained eluate;
s4: the lyophilized sample was redissolved in 100. mu.L of 25 mmol. multidot.L-1To the sodium bicarbonate solution, PNGaseF was added and incubated overnight at 37 ℃;
s5: freeze-drying the solution again and then carrying out nano-LC-MS/MS analysis;
in the above pretreatment process, the buffer solution is ACN-H2O-TFA, 92: 7.9: 0.1, v/v/v; the eluent is ACN-H2O，30：70，v/v。
The invention has the beneficial effects that:
the mHN/Au-GSH aggregates the one-dimensional structural characteristics of the nanofiber, the excellent hydrophilicity of glutathione and Fe3O4The strong magnetic response capability of the glycopeptide probe is integrated, and the glycopeptide probe has the excellent characteristics of high selectivity, high detection sensitivity, strong binding capability, good recovery efficiency and the like in the aspect of glycopeptide enrichment. Furthermore, the practical potential of mHN/Au-GSH nanofibers for glycoproteome analysis was demonstrated by enrichment of low abundance glycopeptides from human serum. The work provides a way for developing new hydrophilic nano-fibers and application thereof in the biomedical field;
the mHN/Au-GSH nanofiber has high glycopeptide detection sensitivity (2fmol), good recovery rate (89.65%), strong binding capacity (100mg/g) and high enrichment selectivity (1: 100).
Drawings
FIG. 1 is a synthesis process diagram of mHN/Au-GSH nano-fiber.
Fig. 2 is a TEM micrograph of HN (1).
Figure 3 is a TEM micrograph of HN (2).
Fig. 4 is a TEM micrograph (1) of mHN.
Fig. 5 is a TEM micrograph (2) of mHN.
FIG. 6 is a TEM micrograph of mHN/Au nanofibers (1).
FIG. 7 is a TEM micrograph of mHN/Au nanofibers (2).
FIG. 8 is an Energy Dispersive Spectroscopy (EDS) elemental distribution plot of the mHN/Au nanofibers prepared.
FIG. 9 is a graph of Fourier Transform Infrared (FTIR) spectral analysis results; wherein, (i) mHN/Au, (ii) mHN/Au-GSH, and (iii) GSH.
FIG. 10 is a diagram showing the result of XRD analysis; wherein, (i) HN, (ii) mHN, and (iii) mHN/Au.
FIG. 11 is a hysteresis loop curve of mHN and mHN/Au-GSH; wherein, (i) mHN, (ii) mHN/Au-GSH.
FIG. 12 is a MALDI-TOFMS spectrum of a direct analysis of a 0.4pmol human IgG digest.
FIG. 13 is a MALDI-TOFMS spectrum of mHN/Au-GSH enriched nanofibers.
FIG. 14 is a MALDI-TOFMS spectrum of mHN-GSH-enriched nanofibers.
FIG. 15 is a MALDI-TOFMS spectrum of HN/Au-GSH enriched nanofibers.
FIG. 16 is a MALDI-TOFMS spectrum enriched with commercial zwitterionic hydrophilic material.
FIG. 17 is a MALDI-TOFMS spectrum enriched by mHN/Au-GSH nanofibers and deglycosylated by PNGaseF.
FIG. 18 is a MALDI-TOF MS spectrum of a 0.25pmol chicken avidin digest direct analysis.
FIG. 19 is a MALDI-TOFMS spectrum enriched with mHN/Au-GSH nanofibers.
FIG. 20 is a MALDI-TOFMS spectrum of mHN/Au-GSH nanofiber enrichment after digestion of 20mmol human IgG.
FIG. 21 is a MALDI-TOFMS spectrum of mHN/Au-GSH nanofiber enrichment after 2mfmol human IgG digestion.
FIG. 22 is a MALDI-TOFMS spectrum of 20fmol human IgG after digestion and enrichment with commercial zwitterionic hydrophilic material.
FIG. 23 is a MALDI-TOFMS spectrum of 2fmol human IgG after digestion and enrichment with commercial zwitterionic hydrophilic material.
FIG. 24 is a MALDI-TOFMS spectrum of a peptide mixture containing a digest of human IgG and a digest of BSA analyzed directly.
FIG. 25 is a MALDI-TOFMS spectrum of a peptide mixture containing a digest of human IgG and a digest of BSA after enrichment with mHN/Au-GSH nanofibers.
Fig. 26 is a graph in which 1: MALDI-TOFMS spectrum of peptide mixture containing human IgG digest and BSA digest after enrichment of Au-GSH nanofibers at a mass ratio of mHN/100. In fig. 12-26, the glycopeptide peaks are labeled with the symbol "×".
Detailed Description
In order to make the technical solution of the present invention clearer and more clear, the present invention is further described below, and any solution obtained by substituting technical features of the technical solution of the present invention with equivalents and performing conventional reasoning falls within the scope of the present invention.
A magnetic nanofiber-based zwitterion hydrophilic material for selectively capturing and identifying glycopeptides utilizes hydroxyapatite nanofibers as a carrier to fix Fe3O4Nanoparticles and Au nanoparticles, via the thiol group of the zwitterionic tripeptide L-glutathione with Au and Fe3O4The surface modification is carried out through the affinity interaction between the two to form mHN/Au-GSH nano-fiber.
Further, the preparation method of the mHN/Au-GSH nano fiber comprises the following steps:
s1: dispersing 2.0g of hydroxyapatite nanofiber and 0.75g of ferric acetylacetonate in 400mL of triethylene glycol to obtain a mixture;
s2: the mixture was transferred to a 1L autoclave and reacted at 220 ℃ for 12 hours, washed with water and ethanol to give fixed Fe3O4A nanofiber product of nanoparticles mHN;
s3: 0.5g of prepared mHN and 25mL of tetrachloroauric acid trihydrate were added to 1L of deionized water to obtain a suspension;
s4: the suspension was stirred at 25 ℃ for 12 hours, 50mg of sodium borohydride was added and stirred for 30 minutes to obtain simultaneous fixed Fe3O4Nanofiber product mHN/Au of nanoparticles and Au nanoparticles;
s5: after washing with deionized water, 0.5g of the HN/Au product was dispersed in an aqueous solution containing 2g of glutathione; stirring at 50 deg.C for 10 hr to obtain mHN/Au-GSH nanofiber.
Further, the concentration of tetrachloroauric acid trihydrate in S3 was 2 mg/mL.
Further, mHN/Au-GSH nanofiber is used for enrichment and identification of glycopeptide.
Furthermore, the HN/Au-GSH nano-fiber can be used for pretreatment before ionization time-of-flight mass spectrometry analysis of human immunoglobulin, chicken avidin protein or bovine serum albumin; the method comprises the following specific steps:
s1: digesting human immunoglobulin, chicken avidin or bovine serum albumin with trypsin;
s2: 400 u g mHN/Au-GSH nanofibers and 2.5 u g human immunoglobulin digests, chicken avidin digests or bovine serum albumin digests into 400 u L buffer solution, at 25 degrees C temperature in 15 minutes;
s3: magnetically separating the material, washing with 2 × 400 μ L buffer solution, and adding 20 μ L eluent to elute and capture glycopeptide;
s4: depositing 1 μ L of the eluate and 1 μ L of DHB on a MALDI plate, and performing MALDI-TOFMS analysis;
in the above pretreatment process, the buffer solution is ACN-H2O-TFA, 92: 7.9: 0.1, v/v/v; eluentIs ACN-H2O, 30: 70, v/v; the concentration of DHB was 25 mg/mL.
Further, the mHN/Au-GSH nanofiber can be used for pretreatment before nano-LC-MS/MS analysis of human serum; the method comprises the following specific steps:
s1: trypsinizing human serum;
s2: 2mg of mHN/Au-GSH nanofibers were incubated with 5. mu.L of human serum digest in 1mL of buffer for 30 min;
s3: adding 2 × 100 μ L of the eluate, and lyophilizing the obtained eluate;
s4: the lyophilized sample was redissolved in 100. mu.L of 25 mmol. multidot.L-1To the sodium bicarbonate solution, PNGaseF was added and incubated overnight at 37 ℃;
s5: freeze-drying the solution again and then carrying out nano-LC-MS/MS analysis;
in the above pretreatment process, the buffer solution is ACN-H2O-TFA, 92: 7.9: 0.1, v/v/v; the eluent is ACN-H2O，30：70，v/v。
For further understanding of the present invention, the following embodiments are provided to illustrate the technical solutions of the present invention in detail, and the scope of the present invention is not limited by the following embodiments.
mHN/characterization of Au-GSH nanofibers
The synthesis of magnetic nanofiber-based zwitterionic hydrophilic materials (mHN/Au-GSH nanofibers) is given in FIG. 1. Briefly, one-dimensional Hydroxyapatite Nanofiber (HN) is synthesized by a solvothermal reaction method and used as modified Fe3O4Nanoparticles and Au nanoparticles. Subsequently, sulfhydryl groups in Glutathione (GSH) were reacted with Au and Fe3O4The nanoparticles react to allow glutathione to be immobilized on the magnetic nanofibers. The mHN/Au-GSH nanofiber has the characteristics of nanofiber, magnetic response capability and hydrophilic glutathione group, and can be used as an adsorbent to specifically capture glycopeptides in the sample preparation process of mass spectrometry glycopeptide omics research.
FIGS. 2, 3, 4, 5, 6 and 7 show the shapes of HN, mHN and mHN/Au nanofibers producedState. HN consists of hydroxyapatite nanowires with pronounced one-dimensional features, including nanometer-sized diameters of 10-20nm, high aspect ratio, and self-assembly into nanofibers (fig. 2, fig. 3). Compared to the smooth surface of HN, a large number of small nanoparticles with an average diameter of about 12nm were decorated at mHN (fig. 4, fig. 5). In addition, after the Au nanoparticles are fixed on mHN, more small nanoparticles appear on the MHN surface, and mHN/Au nanofibers are formed. The average particle size of the modified gold nanoparticles was calculated to be about 11.6nm using the Scherrer equation according to the X-ray diffraction (XRD) analysis results. In addition, the spatial element distribution of P, Ca, Fe and Au from mHN/Au nanofibers was studied using Energy Dispersive Spectroscopy (EDS) elemental mapping. As shown in fig. 8, the four element signals are uniformly localized, further confirming the signal peaks, revealing Fe3O4And successful immobilization of Au nanoparticles in mHN/Au nanofibers.
Modification of glutathione was confirmed by Fourier Transform Infrared (FTIR) spectroscopic analysis. In FIG. 9, a hydroxyl group (3573 cm)-1)，PO4 3-Radicals (1093, 1031, 962, 605 and 565cm-1) Absorption peaks of (2), adsorbed water was observed in the infrared spectrum of mHN/Au (3436 and 1633 cm)-1)。565cm-1Absorption peak and Fe3O4(560cm-1) Overlap of characteristic absorption peaks of (a). In addition, fingerprint peaks belonging to glutathione were clearly found in fig. 9. These results demonstrate successful glutathione immobilization in Fe3O4And Au nanofibers.
In addition, the structure was also analyzed by XRD. In fig. 10, the characteristic peak of HN is consistent with the analysis result of hydroxyapatite. In contrast, the relative increase in intensity of the diffraction peak marked with the black symbol' at 35.5 ° is due to Fe in the mHNX ray diffraction pattern3O4Due to the magnetic component of (a). In addition, in the XRD pattern of mHN/Au nanofiber, a new peak with black symbol '#' at 38.3 ℃ belongs to Au.
As shown in FIG. 11, hysteresis loops of mHN and mHN/Au-GSH were determined, indicating that these two materials have a supermagnetic characteristic. The saturation magnetization values of mHN and mHN/Au-GSH were found to be 6, respectively.76 and 5.51emu g-1. Although the final nanofibers have a relatively low saturation magnetization value, it is sufficient to allow rapid magnetic separation of the nanofibers from aqueous solutions.
Glycopeptide enrichment performance of mHN/Au-GSH nanofiber measured by standard glycoprotein digestion method
Human immunoglobulin IgG trypsin digest was used as a model sample to study the glycopeptide enrichment performance of the prepared mHN/Au-GSH nanofibers. As shown in fig. 12, no glycopeptide peaks were observed in the non-enriched human IgG digest. However, after enrichment with mHN/Au-GSH nanofibers, the glycopeptide peak with strong signal intensity dominates the mass spectrum and there is hardly any interference from non-glycopeptide peaks (FIG. 13). We performed a total of 21 glycopeptides for detection and identification. In addition, mHN-GSH nanofibers, non-magnetic nanofibers (HN/Au-GSH), and commercial ZIC-HILIC materials were used as controls. In fig. 14, 15 and 16, only 17, 20 and 18 glycopeptide peaks were detected, respectively. After mHN/Au-GSH nanofiber enrichment, the identified glycopeptide peak has signal intensity and signal-to-noise ratio (S/N) which are obviously higher than those of a control material after treatment. For example, glycopeptide peaks enriched with mHN/Au-GSH, mHN-GSH, HN/Au-GSH, and commercial ZIC-HILIC materials have S/N ratios of 810.7, 464.7, 564.5, and 492.6 at a mass to charge ratio of 2764.1, respectively. The result shows that the enrichment performance of mHN/Au-GSH is superior to that of mHN-GSH, HN/Au-GSH and commercial ZIC-HILIC materials. In addition, the captured glycopeptide is deglycosylated by PNGaseF (glycoamidase). In FIG. 17, all the glycopeptide peaks detected disappeared, leaving only the two major deamidated peptides (EEQFN # STFR, EEQYN # STYR), indicating that all the peaks observed in FIG. 13 are related to N-bond glycopeptides. The unique one-dimensional structure, abundant zwitterion groups and strong magnetic responsiveness contribute to providing excellent enrichment performance. These results all confirm that the prepared mHN/Au-GSH nanofiber has high enrichment specificity and efficiency on glycopeptide.
In addition, a trypsin digest of chicken avidin was also used. Mass spectra of chicken avidin digests As shown in FIG. 18, 8 glycopeptide peaks with low signal intensity and S/N ratio were observed due to the simultaneous presence of a large number of non-glycopeptides. Importantly, after the selective enrichment process using mHN/Au-GSH nanofibers, the 14 glycopeptide peaks in the mass spectrum (FIG. 19) were affected with improved signal intensity and S/N ratio. These analysis results demonstrate that the prepared mHN/Au-GSH nanofibers have high selectivity and efficiency for general glycopeptide enrichment.
The detection sensitivity of the prepared mHN/Au-GSH nanofiber to glycopeptide is determined. As shown in fig. 20, 12 glycopeptide peaks were clearly observed in a 20fmol human immunoglobulin IgG digest after the enrichment process. When the amount of human IgG digestion was as low as 2fmol (FIG. 21), four glycopeptide peaks were still observed, with a maximum S/N ratio of 22.8. The detection sensitivity of mHN/Au-GSH nanofiber for glycopeptide was considered to be 2 fmol. For comparison, the detection sensitivity of commercial hydrophilic materials was also tested. When the amount of human IgG digestion was 20fmol and enriched with commercial hydrophilic material, only 8 glycopeptide peaks could be detected (fig. 22). On the other hand, when the amount of human IgG digested was only 2fmol (FIG. 23), no glycopeptide peak was observed. The mHN/Au-GSH nanofiber studied at this time has higher detection sensitivity on abundant low-abundance glycopeptides, and the performance of the nanofiber is superior to that of CS @ PGMA @ IDA-Ti4+(45fmol)、GO-Fe3O4/SiO2AuNWs/L-cysteine (10fmol), MoS2Au-NP-L-cysteine (10fmol) and Fe3O4@ TpPa are several hydrophilic materials.
According to the previous literature, the recovery of enrichment of 6 targeted glycopeptides released from human IgG was evaluated by isotopic dimethyl labelling. The average enrichment recovery achieved was 89.65%. Compared with Resin @ Polysaccharide (73%), GO/Fe which are reported in the past3O4/Au/PEG(59.2％)、Fe3O4@MPS@PMAC(82％)、MIL-101(NH2) Compared with several hydrophilic materials of @ Au-Cysteine (80%) and Resin/Au-Cysteine (79%), the mHN/Au-GSH nanofiber prepared by the method has better capture capability.
At the same time, the binding capacity of the confirmed mHN/Au-GSH nanofiber is measured. A fixed mass of human IgG digest (3. mu.g) was incubated with a mass of mHN/Au-GSH. After a period of time, the detection is carried out when mHN/Au-GSH is usedAt 30. mu.g, the signal intensities of the six selected glycopeptides reached a maximum. mHN/Au-GSH nano-fiber with glycopeptide binding capacity of 100mg g-1. This is probably due to the strong multivalent hydrophilic interaction between the glycopeptide and the MHN/Au-GSH nanofibers due to the abundance of zwitterionic hydrophilic groups. The experiment result shows that the combination capability of the MHN/Au-GSH nano-fiber is stronger than that of Fe3O4@CSMCNCs(17.5mg/g)、Fe3O4-DA-Maltose (43mg/g) and Fe3O4@PGMA@Au-L-cysteine(75mg/g)。
The enrichment selectivity of mHN/Au-GSH nanofibers was further investigated. As shown in fig. 24, by directly analyzing the polypeptide mixture containing the human IgG digest and the BSA digest, we found that the non-glycopeptide peak was dominant in the mass spectrum and that the glycopeptide peak was not clearly identified. However, after enrichment with mHN/Au-GSH nanofibers, the non-glycopeptides disappeared significantly, and the signal intensities of 19 and 17 glycopeptides detected were significantly improved (FIGS. 25 and 26), which provides more evidence for enrichment of glycopeptides with mHN/Au-GSH nanofibers.
Selective enrichment and identification of glycopeptides in human serum
Human serum has attracted considerable attention as a typical clinical specimen, a potential biomarker for diagnosing diseases. The advantages of mHN/Au-GSH nanofiber in glycopeptide enrichment prove that human serum glycoprotein is analyzed according to the scheme of proteolysis, mHN/Au-GSH nanofiber glycopeptide enrichment, PNGase F deglycosylation and mass spectrum identification. As a result, a total of 246 glycopeptides of 104 glycoproteins were identified by only three independent analyses of 1. mu.L human serum. In contrast, commercial zwitterionic hydrophilic materials identified only 162 glycopeptides from 69 glycoproteins. The structural differences of these affinity materials contribute to their diverse abilities in glycopeptide enrichment. These results indicate that the prepared mHN/Au-GSH nanofiber has high specificity and high efficiency in capturing low-abundance glycopeptides, and has great potential in researching the glycoproteome of complex biological samples.
The experimental procedures in this example are conventional, unless otherwise specified. The experimental materials used in the present examples were all purchased from the market unless otherwise specified.
Transmission Electron Microscope (TEM) images and Energy Dispersive Spectroscopy (EDS) elemental mapping were obtained with a transmission electron microscope (feitecnag 2F20, USA). The infrared spectra were obtained using a Thermo Nicolet380 Fourier transform infrared spectrometer (Nicolet, Wisconsin, USA). X-ray diffractometry (RigakuD/max2550V, CuK alpha radiation,
) An X-ray powder diffraction (XRD) pattern was recorded. The saturation magnetization curve was tested on a physical property measurement system (PPMS, USA) at room temperature. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) experiments were performed in linear positive mode on an AB Sciex5800 mass spectrometer (ABSciex, CA) using a Nd/YAG laser at 355 nm.
Claims (2)
1. The magnetic nanofiber-based zwitterion hydrophilic material for selectively capturing and identifying glycopeptides is characterized in that the magnetic nanofiber-based zwitterion hydrophilic material is prepared by fixing Fe by taking hydroxyapatite nanofiber as a carrier3O4Nanoparticles and Au nanoparticles, via the thiol group of the zwitterionic tripeptide L-glutathione with Au and Fe3O4The surface modification is carried out by the affinity interaction, and the specific preparation method comprises the following steps:
s1: dispersing 2.0g of hydroxyapatite nanofiber and 0.75g of ferric acetylacetonate in 400mL of triethylene glycol to obtain a mixture;
s2: the mixture was transferred to a 1L autoclave and reacted at 220 ℃ for 12 hours, washed with water and ethanol to give fixed Fe3O4A nanofiber product of nanoparticles mHN;
s3: 0.5g of prepared mHN and 25mL of tetrachloroauric acid trihydrate were added to 1L of deionized water to obtain a suspension;
s4: the suspension was stirred at 25 ℃ for 12 hours, 50mg of sodium borohydride was added and stirred for 30 minutes to obtain simultaneous fixed Fe3O4Nanofiber product mH of nanoparticles and Au nanoparticlesN/Au；
S5: after washing with deionized water, 0.5g of the mHN/Au product was dispersed in an aqueous solution containing 2g of the zwitterionic tripeptide L-glutathione; stirring for 10 hours at 50 ℃ to obtain the magnetic nanofiber-based zwitterionic hydrophilic material.
2. The magnetic nanofiber-based zwitterionic hydrophilic material for selectively capturing and recognizing a glycopeptide according to claim 1, wherein the concentration of tetrachloroaurate trihydrate in S3 is 2 mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910340139.3A CN110130099B (en) | 2019-04-25 | 2019-04-25 | Magnetic nanofiber-based zwitterionic hydrophilic materials for selective capture and recognition of glycopeptides |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910340139.3A CN110130099B (en) | 2019-04-25 | 2019-04-25 | Magnetic nanofiber-based zwitterionic hydrophilic materials for selective capture and recognition of glycopeptides |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110130099A CN110130099A (en) | 2019-08-16 |
| CN110130099B true CN110130099B (en) | 2021-06-08 |
Family
ID=67571212
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910340139.3A Expired - Fee Related CN110130099B (en) | 2019-04-25 | 2019-04-25 | Magnetic nanofiber-based zwitterionic hydrophilic materials for selective capture and recognition of glycopeptides |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110130099B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111841511A (en) * | 2020-07-13 | 2020-10-30 | 浙江农林大学 | Magnetic nanoparticle-graphene oxide nanocomposite and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101723311A (en) * | 2009-11-06 | 2010-06-09 | 武汉理工大学 | Magnetic flourescent nano material for magnetic-photo sensor and preparation method thereof |
| CN102431986A (en) * | 2011-10-08 | 2012-05-02 | 山东大学 | Method for preparing hydroxyapatite nano fiber |
| CN105823847A (en) * | 2016-03-16 | 2016-08-03 | 复旦大学 | Glycopeptide enriching and detecting method of amphoteric hydrophilic composite nano material |
| CN105862391A (en) * | 2016-03-29 | 2016-08-17 | 苏州大学 | Preparation method of BC@metal oxide composite nanofiber |
| CN107083241A (en) * | 2017-06-19 | 2017-08-22 | 中南大学 | Water soluble hydroxy apatite fluorescent nano-particle and its preparation and activity and the method for inhibition for testing and analyzing PKA |
| CN107486177A (en) * | 2017-09-15 | 2017-12-19 | 四川大学 | Magnetic metal organic framework nanosphere with how affine site and preparation method and application |
-
2019
- 2019-04-25 CN CN201910340139.3A patent/CN110130099B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101723311A (en) * | 2009-11-06 | 2010-06-09 | 武汉理工大学 | Magnetic flourescent nano material for magnetic-photo sensor and preparation method thereof |
| CN102431986A (en) * | 2011-10-08 | 2012-05-02 | 山东大学 | Method for preparing hydroxyapatite nano fiber |
| CN105823847A (en) * | 2016-03-16 | 2016-08-03 | 复旦大学 | Glycopeptide enriching and detecting method of amphoteric hydrophilic composite nano material |
| CN105862391A (en) * | 2016-03-29 | 2016-08-17 | 苏州大学 | Preparation method of BC@metal oxide composite nanofiber |
| CN107083241A (en) * | 2017-06-19 | 2017-08-22 | 中南大学 | Water soluble hydroxy apatite fluorescent nano-particle and its preparation and activity and the method for inhibition for testing and analyzing PKA |
| CN107486177A (en) * | 2017-09-15 | 2017-12-19 | 四川大学 | Magnetic metal organic framework nanosphere with how affine site and preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110130099A (en) | 2019-08-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Facile synthesis of zwitterionic polymer-coated core–shell magnetic nanoparticles for highly specific capture of N-linked glycopeptides | |
| Wang et al. | Facile synthesis of magnetic covalent organic frameworks for the hydrophilic enrichment of N-glycopeptides | |
| Yao et al. | One-step functionalization of magnetic nanoparticles with 4-mercaptophenylboronic acid for a highly efficient analysis of N-glycopeptides | |
| Huan et al. | A magnetic nanofiber-based zwitterionic hydrophilic material for the selective capture and identification of glycopeptides | |
| Jiao et al. | Ultrathin Au nanowires assisted magnetic graphene-silica ZIC-HILIC composites for highly specific enrichment of N-linked glycopeptides | |
| Kailasa et al. | Nanomaterial-based miniaturized extraction and preconcentration techniques coupled to matrix-assisted laser desorption/ionization mass spectrometry for assaying biomolecules | |
| Pan et al. | Dual-functionalized magnetic bimetallic metal-organic framework composite for highly specific enrichments of phosphopeptides and glycopeptides | |
| Wu et al. | Thiol-ene click synthesis of L-cysteine-bonded zwitterionic hydrophilic magnetic nanoparticles for selective and efficient enrichment of glycopeptides | |
| Xu et al. | Boronic acid-functionalized detonation nanodiamond for specific enrichment of glycopeptides in glycoproteome analysis | |
| Zhao et al. | Synthesis of magnetic zwitterionic–hydrophilic material for the selective enrichment of N-linked glycopeptides | |
| Zheng et al. | Click synthesis of glucose-functionalized hydrophilic magnetic mesoporous nanoparticles for highly selective enrichment of glycopeptides and glycans | |
| Zhao et al. | Recent advances in the application of core–shell structured magnetic materials for the separation and enrichment of proteins and peptides | |
| Feng et al. | Facile and easily popularized synthesis of L-cysteine-functionalized magnetic nanoparticles based on one-step functionalization for highly efficient enrichment of glycopeptides | |
| Zhang et al. | Phenylboronic acid functionalized C3N4 facultative hydrophilic materials for enhanced enrichment of glycopeptides | |
| Ma et al. | Ligand-free strategy for ultrafast and highly selective enrichment of glycopeptides using Ag-coated magnetic nanoarchitectures | |
| CN107727480B (en) | Ultrathin lamellar functionalized molybdenum disulfide nano composite material and application thereof in glycopeptide enrichment | |
| Long et al. | Low-cost iron oxide magnetic nanoclusters affinity probe for the enrichment of endogenous phosphopeptides in human saliva | |
| Chu et al. | One-pot preparation of hydrophilic citric acid-magnetic nanoparticles for identification of glycopeptides in human saliva | |
| CN106732474B (en) | Magnetic nano material, preparation method thereof and application of magnetic nano material in enrichment analysis of glycosylated peptide segments | |
| Li et al. | Preparation of titanium-grafted magnetic mesoporous silica for the enrichment of endogenous serum phosphopeptides | |
| CN110779789A (en) | Preparation of hydrophilic group modified two-dimensional magnetic nano material and application of hydrophilic group modified two-dimensional magnetic nano material in large-scale enrichment of glycopeptide | |
| Chen et al. | Preparation of C60‐functionalized magnetic silica microspheres for the enrichment of low‐concentration peptides and proteins for MALDI‐TOF MS analysis | |
| Pu et al. | Elution-free ultra-sensitive enrichment for glycopeptides analyses: Using a degradable, post-modified Ce-metal–organic framework | |
| Zhang et al. | L-cysteine functionalized straticulate C3N4 for the selective enrichment of glycopeptides | |
| CN108906007A (en) | A kind of preparation method and applications of the hydrophilic magnetic composite microballoon of glycosyl |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210608 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |