CN219897676U - Glycopeptide enrichment device - Google Patents
Glycopeptide enrichment device Download PDFInfo
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- CN219897676U CN219897676U CN202321524724.7U CN202321524724U CN219897676U CN 219897676 U CN219897676 U CN 219897676U CN 202321524724 U CN202321524724 U CN 202321524724U CN 219897676 U CN219897676 U CN 219897676U
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- pipe
- tube
- liner
- glycopeptide
- enrichment device
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- 102000002068 Glycopeptides Human genes 0.000 title claims abstract description 82
- 108010015899 Glycopeptides Proteins 0.000 title claims abstract description 82
- 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 title claims abstract description 75
- 239000000945 filler Substances 0.000 claims abstract description 31
- 239000012528 membrane Substances 0.000 claims abstract description 7
- 238000007789 sealing Methods 0.000 claims description 14
- 229920000742 Cotton Polymers 0.000 claims description 5
- 238000002013 hydrophilic interaction chromatography Methods 0.000 claims 1
- 238000000034 method Methods 0.000 description 15
- 238000011033 desalting Methods 0.000 description 7
- 102000007079 Peptide Fragments Human genes 0.000 description 6
- 108010033276 Peptide Fragments Proteins 0.000 description 6
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 238000000926 separation method Methods 0.000 description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000010612 desalination reaction Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 125000005842 heteroatom Chemical group 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 239000002120 nanofilm Substances 0.000 description 2
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 229920006122 polyamide resin Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical group 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000003749 cleanliness Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Abstract
The utility model provides a glycopeptide enrichment device, which comprises a plurality of liner tubes and a molecular section membrane, wherein the liner tubes are of circular tube structures, a first connecting part and a second connecting part are respectively arranged at two ends of each liner tube, the liner tubes are coaxially arranged, and two adjacent liner tubes are communicated through the first connecting part and the second connecting part; at least one of the lining pipes is fixedly provided with the molecular cut-off film which is used for bearing the enrichment filler of glycopeptides. According to the utility model, through arranging a plurality of liner tubes and selectively arranging the molecular cut-off films in the liner tubes, one or more glycopeptide enrichment filler can be loaded in the glycopeptide enrichment device according to actual needs, so that the enrichment rate of the glycopeptide enrichment device is improved.
Description
Technical Field
The utility model relates to the technical field of separation and purification of glycopeptides, in particular to a glycopeptides enrichment device.
Background
In recent years, researchers have been working on developing strategies for enriching and separating glycopeptides to facilitate and improve glycosylation analysis in complex samples, and various glycopeptide enrichment methods such as immunoaffinity chromatography, lectin affinity method, hydrazide method, titanium dioxide (TiO 2) enrichment method, hydrophilic interaction liquid chromatography (HILIC) method, porous Graphitized Carbon (PGC) method, polyamide resin (DPA) method, etc. have been developed.
The utility model of the prior publication No. CN212845217U discloses a device capable of combining an HILIC chromatographic column with polypeptide desalting, combines HILIC enriched glycopeptide with polypeptide desalting, adjusts the proportion of an organic phase of eluent through different hydrophilic actions of HILIC and C18, so that the glycopeptide is enriched in the HILIC column material first, then eluted into the C18 desalting column, combines enrichment and desalting, reduces the operation flow, improves the efficiency, solves the problems of complicated operation, long time consumption and the like of the existing glycopeptide enrichment and desalting, has a simple structure and convenient operation, and is suitable for popularization and application.
In the above technical scheme, in order to improve the efficiency of the glycopeptide enrichment operation, the HILIC enrichment glycopeptide and polypeptide desalting are combined, but because the glycopeptide of a complex sample is difficult to be enriched in a high-efficiency and broad-spectrum manner by a single glycopeptide enrichment mode, the glycopeptide loss of the glycopeptide enrichment device is more, and the glycopeptide enrichment rate is reduced.
Disclosure of Invention
In view of the above, the utility model provides a glycopeptide enrichment device, which can combine various glycopeptide enrichment modes together to improve the enrichment rate of glycopeptides.
The technical scheme of the utility model is realized as follows: the utility model provides a glycopeptide enrichment device, which comprises a plurality of lining pipes and molecular cut-off films, wherein,
the lining pipes are of circular pipe structures, a first connecting part and a second connecting part are respectively arranged at two ends of the lining pipes, a plurality of the lining pipes are coaxially arranged, and two adjacent lining pipes are communicated through the first connecting part and the second connecting part;
at least one of the lining pipes is fixedly provided with the molecular cut-off film which is used for bearing the enrichment filler of glycopeptides.
On the basis of the above technical solution, preferably, the first connection portion is a circular groove structure, and is opened at one end of the liner tube.
Still further preferably, the second connecting portion is a circular ring-shaped bulge-like structure, and the second connecting portion is integrally formed at one end of the liner tube away from the first connecting portion.
Still more preferably, a top tube and a cover are also included, wherein,
the jacking pipe is detachably fixed on the liner pipe positioned at one end of the glycopeptide enrichment device;
the sealing cover is arranged at one end of the jacking pipe far away from the liner pipe in a sealing way and is used for selectively sealing the jacking pipe.
Still further preferably, the apparatus further comprises a bottom tube detachably fixed to the liner tube at an end of the glycopeptide enrichment apparatus remote from the top tube, and the end of the bottom tube remote from the liner tube is tapered.
Still further preferably, the first connecting portion is disposed at an end of the liner tube near the bottom tube;
the second connecting part is arranged at one end of the liner tube, which is close to the jacking pipe.
Still more preferably, the first connecting portion is provided at one end of the top pipe close to the liner pipe;
the bottom pipe is close to one end of the liner pipe and is integrally formed with the second connecting part.
On the basis of the technical scheme, preferably, the top pipe and the bottom pipe are fixedly provided with one molecular section membrane.
Still more preferably, two of the liners are provided, and each of the liners is fixedly provided with one molecular cut-off film, and the four molecular cut-off films are respectively used for bearing C18 filler, HILIC filler, PGC filler and DPA filler, and absorbent cotton is filled in the bottom pipe.
Compared with the prior art, the glycopeptide enrichment device has the following beneficial effects:
(1) By arranging a plurality of liner tubes and selectively arranging molecular cut-off films in the liner tubes, one or more glycopeptide enrichment filler can be loaded in the glycopeptide enrichment device according to actual needs, so that the enrichment rate of the glycopeptide enrichment device is improved;
(2) The top pipe and the sealing cover are arranged, so that the glycopeptide enrichment device can be covered and plugged, and the bottom pipe with one conical end is arranged, so that the glycopeptide enrichment device can be matched with other equipment conveniently, and the glycopeptide enrichment device can be used conveniently;
(3) Through set up first connecting portion and second connecting portion on bushing pipe, top tube and bottom tube, not only can conveniently carry out the free equipment to this glycopeptide enrichment device, still promoted the sealing performance of this glycopeptide enrichment device.
Drawings
In order to more clearly illustrate the embodiments of the utility model or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the utility model, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a perspective view of a glycopeptide enrichment device of the present utility model;
FIG. 2 is a perspective view of a glycopeptide enrichment device of the present utility model;
FIG. 3 is a perspective view of a glycopeptide enrichment device of the present utility model;
FIG. 4 is a perspective view of a glycopeptide enrichment device of the present utility model.
Wherein: 1. a liner tube; 11. a first connection portion; 12. a second connecting portion; 2. molecular membrane cutting; 3. jacking pipes; 4. a cover; 5. a bottom tube.
Detailed Description
The following description of the embodiments of the present utility model will clearly and fully describe the technical aspects of the embodiments of the present utility model, and it is apparent that the described embodiments are only some embodiments of the present utility model, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present utility model without making any inventive effort, are intended to fall within the scope of the present utility model.
As shown in fig. 1 to 4, a glycopeptide enrichment device of the present utility model comprises a plurality of liners 1 and a molecular cut-off membrane 2.
The liner tube 1 is used for circulating a glycopeptide sample, the liner tube 1 is of a circular tube structure, a first connecting portion 11 and a second connecting portion 12 are respectively arranged at two ends of the liner tube 1, a plurality of liner tubes 1 are coaxially arranged, and two adjacent liner tubes 1 are communicated through the first connecting portion 11 and the second connecting portion 12.
The molecular cut-off film 2 is used for bearing the enrichment filler of the glycopeptide, the molecular cut-off film 2 is fixedly arranged in at least one liner tube 1, namely, one molecular cut-off film 2 can be arranged in a plurality of liner tubes 1, so that the liner tube 1 is filled with one glycopeptide enrichment filler, the glycopeptide is enriched in an enrichment mode, of course, the plurality of liner tubes 1 can also be provided with a plurality of molecular cut-off films 2, so that two or more glycopeptide enrichment fillers can be filled in the liner tube 1, and the multiple enrichment modes are compounded in the glycopeptide enrichment device, and the glycopeptide enrichment efficiency of the glycopeptide enrichment device is higher and the content of the heteropolypeptide is lower by combining the advantages of different glycopeptide enrichment modes; in addition, the number of the liner tubes 1 can be selected according to actual needs, and the number of the liner tubes 1 provided with the molecular cut-off films 2 can be selected according to actual needs, so that the glycopeptide enrichment device can be freely assembled.
As a preferred embodiment, the first connection portion 11 should be provided as a circular groove-like structure, which is opened at one end of the liner tube 1; correspondingly, the second connecting part 12 is preferably provided with a circular convex structure, the second connecting part 12 is integrally formed at one end of the liner tube 1 far away from the first connecting part 11, as shown in fig. 2, and the connection and fixation of the adjacent liner tubes 1 can be realized by utilizing the clamping connection between the second connecting part 12 and the first connecting part 11; the connecting structure is simple, easy to process and convenient for free assembly and sealing connection of the glycopeptide enrichment device.
In order to ensure the cleanliness of the enrichment operation, a jacking pipe 3 and a sealing cover 4 can be further arranged, the jacking pipe 3 is detachably fixed on the liner pipe 1 at one end of the glycopeptide enrichment device, and the sealing cover 4 is arranged at one end, far away from the liner pipe 1, of the jacking pipe 3 in a sealing manner and is used for selectively sealing the jacking pipe 3 so as to prevent impurities in the operation environment from entering the liner pipe 1.
Further, in order to facilitate inserting the glycopeptide enrichment device into the waste liquid collecting device, a bottom tube 5 can be further arranged, the bottom tube 5 is detachably fixed on the liner tube 1 positioned at one end, far away from the top tube 3, of the glycopeptide enrichment device, and one end, far away from the liner tube 1, of the bottom tube 5 is conical.
Further, in order to improve the connection tightness between the liners 1, the first connection portion 11 may be disposed at an end of the liner 1 near the bottom pipe 5, and the second connection portion 12 may be disposed at an end of the liner 1 near the top pipe 3; correspondingly, in order to facilitate the fixation of the top pipe 3 and the bottom pipe 5, a first connecting portion 11 may be formed at one end of the top pipe 3 close to the liner pipe 1, and a second connecting portion 12 may be integrally formed at one end of the bottom pipe 5 close to the liner pipe 1.
As a preferred embodiment, a molecular cut-off film 2 is fixedly arranged in each of the top pipe 3 and the bottom pipe 5, and when the number of the molecular cut-off films 2 needs to be adjusted in the enrichment operation process, the liner pipe 1 only needs to be replaced, and the adjustment of the top pipe 3 and the bottom pipe 5 is not needed.
Furthermore, preferably, two liners 1 are arranged, one molecular cut film 2 is fixedly arranged in each liner 1, the molecular cut films 2 in the top pipe 3 and the bottom pipe 5 are added, four molecular cut films 2 are arranged in the glycopeptide enrichment device, the four molecular cut films 2 are respectively used for bearing C18 filler, HILIC filler, PGC filler and DPA filler, absorbent cotton is filled in the bottom pipe 5, the absorbent cotton can enable air inside and outside the device to circulate, and harmful substances such as external bacteria and the like can be prevented from entering the device, so that the protective effect is achieved, as shown in fig. 1, the C18 filler, HILIC filler, PGC filler and DPA filler are sequentially arranged on the four molecular cut films 2 in the glycopeptide enrichment device from top to bottom, and absorbent cotton is further arranged at the bottom in the bottom pipe 5, after enzyme-cut complex samples are added into the device, and on-line desalination is realized through the C18 filler; then through HILIC filler, the peptide fragment of the complex sample is subjected to primary glycosylated peptide fragment enrichment, and a liquid chromatography (HILIC) enrichment method can separate glycoprotein and glycopeptide and glycan derived from glycoprotein, wherein the enrichment method uses a hydrophilic stationary phase and a relatively hydrophobic organic mobile phase, and the glycopeptide can be more firmly combined on the HILIC column due to the influence of a hydrophilic carbohydrate part in the structure, so that the glycopeptide can be separated from non-glycosylated peptide fragment; then through PGC filler, secondary glycosylated peptide enrichment is carried out, the Porous Graphitized Carbon (PGC) method is a high-efficiency material for separation and solid phase extraction of glycan and glycopeptide, and during the separation process by using a solid phase extraction column, the glycopeptide is reserved, but not the glycopeptide flows out, so that the purpose of separation is achieved; finally, through DPA filler, the peptide is subjected to third glycosylation peptide enrichment, and according to a reverse phase mechanism, the polyamide resin (DPA) method adsorbs polar glycopeptide fragments from an aqueous solution or a methanol solution through strong hydrogen bonding between hydroxyl groups of a compound and amide groups of the resin, and the non-glycopeptide fragments flow out; by combining multiple glycopeptide enrichment methods, the selectivity and the sensitivity can be improved, so that the efficiency of glycopeptide separation and enrichment is improved, and the ratio of the hetero peptide fragments is reduced.
The application method of the glycopeptide enrichment device provided by the utility model comprises the following steps:
when the glycopeptide enrichment operation is carried out, firstly, a corresponding number of liner tubes 1 are selected according to actual demands, the enrichment device is assembled, liner tubes 1 without filling materials can be selected from liner tubes 1 without molecular film cutting 2, corresponding molecular film cutting 2 are respectively filled with desalting filler and corresponding enrichment filler, then, the complex sample peptide fragments after enzyme cutting are added into the enrichment device for desalination and enrichment operation, and finally, glycopeptides in a plurality of enrichment fillers are eluted and mixed together, so that glycopeptides with wider enrichment range and lower hetero peptide fragment ratio can be obtained.
The foregoing description of the preferred embodiments of the utility model is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the utility model.
Claims (9)
1. A glycopeptide enrichment device, characterized in that: comprises a plurality of lining pipes (1) and a molecular section film (2), wherein,
the lining pipes (1) are of circular pipe structures, a first connecting part (11) and a second connecting part (12) are respectively arranged at two ends of each lining pipe, a plurality of the lining pipes (1) are coaxially arranged, and two adjacent lining pipes (1) are communicated through the first connecting part (11) and the second connecting part (12);
at least one lining tube (1) is internally fixedly provided with the molecular cut-off film (2) for bearing the enrichment filler of glycopeptides.
2. A glycopeptide enrichment device according to claim 1, wherein: the first connecting part (11) is of a circular groove-shaped structure and is arranged at one end of the liner tube (1).
3. A glycopeptide enrichment device according to claim 2, wherein: the second connecting portion (12) is of a circular-ring-shaped bulge-shaped structure, and the second connecting portion (12) is integrally formed at one end, far away from the first connecting portion (11), of the liner tube (1).
4. A glycopeptide enrichment device according to claim 3, wherein: also comprises a jacking pipe (3) and a sealing cover (4), wherein,
the jacking pipe (3) is detachably fixed on the liner pipe (1) positioned at one end of the glycopeptide enrichment device;
the sealing cover (4) is arranged at one end, far away from the liner tube (1), of the jacking tube (3) in a sealing mode and is used for selectively sealing the jacking tube (3).
5. A glycopeptide enrichment device according to claim 4, wherein: the device is characterized by further comprising a bottom tube (5), wherein the bottom tube (5) is detachably fixed on the liner tube (1) which is positioned at one end of the glycopeptide enrichment device, which is far away from the top tube (3), and one end of the bottom tube (5), which is far away from the liner tube (1), is in a conical shape.
6. A glycopeptide enrichment device according to claim 5, wherein: the first connecting part (11) is arranged at one end of the liner tube (1) close to the bottom tube (5);
the second connecting part (12) is arranged at one end of the liner tube (1) close to the jacking pipe (3).
7. A glycopeptide enrichment device according to claim 6, wherein: one end of the jacking pipe (3) close to the liner pipe (1) is provided with the first connecting part (11);
the second connecting part (12) is integrally formed at one end of the bottom pipe (5) close to the liner pipe (1).
8. A glycopeptide enrichment device according to any of claims 5-7, wherein: the top pipe (3) and the bottom pipe (5) are internally and fixedly provided with the molecular section membrane (2).
9. A glycopeptide enrichment device according to claim 8, wherein: the lining pipe (1) is provided with two, and each lining pipe (1) is internally and fixedly provided with one molecular section membrane (2), and four molecular section membranes (2) are respectively used for bearing C18 filler, HILIC filler, PGC filler and DPA filler, and absorbent cotton is filled in the bottom pipe (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321524724.7U CN219897676U (en) | 2023-06-14 | 2023-06-14 | Glycopeptide enrichment device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321524724.7U CN219897676U (en) | 2023-06-14 | 2023-06-14 | Glycopeptide enrichment device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219897676U true CN219897676U (en) | 2023-10-27 |
Family
ID=88435214
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202321524724.7U Active CN219897676U (en) | 2023-06-14 | 2023-06-14 | Glycopeptide enrichment device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN219897676U (en) |
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2023
- 2023-06-14 CN CN202321524724.7U patent/CN219897676U/en active Active
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