CN106018574A - Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture - Google Patents
Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture Download PDFInfo
- Publication number
- CN106018574A CN106018574A CN201610291376.1A CN201610291376A CN106018574A CN 106018574 A CN106018574 A CN 106018574A CN 201610291376 A CN201610291376 A CN 201610291376A CN 106018574 A CN106018574 A CN 106018574A
- Authority
- CN
- China
- Prior art keywords
- parts
- component
- screw
- conduit
- way
- 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.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6052—Construction of the column body
- G01N30/606—Construction of the column body with fluid access or exit ports
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention provides a fractionation device for carrying out simple and rapid ion exchange on a peptide or protein mixture. The fractionation device is low in manufacturing cost, while can obtain the same fractionation effect as a corresponding commercial product. The device comprises a component A, a component B and a component C. The component B is a chromatographic column, wherein A guide pipe penetrates through the centers of two hollow screws, the guide pipe stretches into a two-way parts on the left and right sides, the left and right ends of the two-way parts are provided with internal threads, the two hollow screws are provided with external threads and are in threaded connection with the left and right two-way parts, and sealing iron loops and screen plates are sequentially arranged in front of the two hollow screws respectively. According to the component A, a guide pipe penetrates through the centers of the two hollow screws, one screw is in threaded connection with a two-way part on the left side of the component B, and a sealing iron loop is arranged in front of the screw of the component A. According to the component C, a hollow screw is in threaded connection with a two-way part in the component B through a sealing iron loop, and then a capillary tube for collecting fraction is inserted into the hollow screw of the component C.
Description
Technical field
The present invention relates to the proteomics of life science, in particular to high-efficient liquid phase chromatogram technology field.
Background technology
The qualitative and quantitative analysis of complicated polypeptide and protein mixture is presently mainly and is completed by this instrument analytical method of Multidimensional HPLC-mass spectrum.Multidimensional HPLC is mainly formed by the Orthogonal Composite such as ion exchange chromatography, reversed phase chromatography；Reversed phase chromatography is compatible because of its elution flow phase and mass spectrum, and has high-resolution feature simultaneously, is typically used for the most one-dimensional being used in conjunction with mass spectrum, it is achieved ON-LINE SEPARATION analysis；Ion exchange chromatography is then generally used for the rough prefractionation of original stock sample.But commercialization Ion-exchange high-performance liquid chromatography system is the most expensive, price is at about 100,000 dollars.
Summary of the invention
For solving aforementioned commercial Ion-exchange high-performance liquid chromatography system price costliness problem, the present invention provides a kind of cost of manufacture low, but can obtain, with corresponding commercially produced product same fractionating effect, polypeptide or protein mixture be carried out simple and fast ion exchange fractionating device, including A parts, B parts and C parts, described B parts are chromatographic column, it is to run through a conduit at two hollow screw centers, conduit puts in a left side, in the two-way on right both sides, two-way is left, there is female thread at right two, two hollow screws have external screw thread, respectively with a left side, right two two-ways are spirally connected, two hollow screw fronts have sealing ferrum lasso successively, sieve plate；Described A parts are also to run through a conduit at two hollow screw centers, and one of them screw is spirally connected with the left side two-way of B parts, and there is sealing ferrum lasso the screw front of A parts；Described C parts are to be spirally connected with the two-way in B parts by sealing ferrum lasso with a hollow screw, then collect fraction capillary tube with one, inject in the hollow screw in C parts.
The conduit diameter of described A parts is less than the conduit of B parts, and described conduit uses duroplasts to make.
It is an advantage of the current invention that
(1) this invention simplifies protein mixture piece-rate system, simple in construction.A pressure sample introduction needle and syringe pump are mixed in the separation of this chromatographic system, it is possible to substitute a sufficiently expensive chromatograph of liquid and complete one-dimensional separation.
(2) system flexibility is high, according to being actually needed in experiment, can change length and the upper any filler oneself needing to use of filling of pillar, be particularly suitable for doing the seminar of filler research.This is the condition not available for business liquid-phase chromatographic column.
(3) additionally, itself cost is the most to one's profit.Compared with the conventional thousands of price up to ten thousand of rustless steel liquid-phase chromatographic column, its advantage is also self-evident.
Accompanying drawing explanation
Fig. 1 is decomposing schematic representation of the present invention
Fig. 2 is product schematic diagram of the present invention
Fig. 3 is B exploded view of the present invention
Fig. 4 is C exploded view of the present invention
Fig. 5 is A exploded view of the present invention
Fig. 6 is the effect schematic diagram of the present invention
Label declaration in figure:
1,5,15,8,12-seals ferrum lasso；2,4,9,11,16-hollow screw；3-A component conduit；10-B component conduit；17-capillary tube；6,14-two-way；7,13-sieve plate.
Detailed description of the invention
Refer to shown in accompanying drawing, including A parts, B parts and C parts, described B parts are chromatographic column, being to run through a conduit 10 at two hollow screws 9,11 center, conduit 10 internal diameter is 0.7 millimeter, and conduit 10 puts in the two-way 6,14 on left and right both sides, there is female thread at the left and right two of two-way 10, two hollow screws have external screw thread 9,11, are spirally connected with left and right two two-ways 6,14 respectively, two hollow screw fronts have successively sealing ferrum lasso 8,12, sieve plate 7,13；Described A parts are also to run through a conduit 3 at two hollow screws 2,4 center, a diameter of 0.36 millimeter of conduit 3, and one of them screw 4 is spirally connected with the left side two-way 6 of B parts, and there is sealing ferrum lasso 1,5 screw 2,4 front of A parts；Described C parts are to be spirally connected with the two-way 14 in B parts by sealing ferrum lasso 15 with a hollow screw 16, then collect fraction capillary tube 14 with one, inject in the hollow screw 16 in C parts.
Described conduit uses duroplasts to make.
Filler in chromatographic column, conduit 10 one end opened is injected, and notices that load wants even compact.
After filler filling is complete, then be connected with conduit 10 through sieve plate open one end, the most then chromatographic column body part makes complete.
One end of homemade chromatographic column is connected upper one section of empty conduit, and the loading for protein mixture is used, as shown in fig. 1 on the left-hand side.Its right-hand member then connects the empty capillary column collection for fraction of one section of suitable length, parts as shown on the right side of Fig. 1.
Experimental procedure:
It is connected to made chromatographic column, on six-way valve interface, be balanced relative to brand-new chromatographic column with the flowing of liquid chromatograph.
(1) loading: inject the standard protein sample of 10ul in quantitative loop, complete loading step by mobile phase A.Repeat this process three times.
(2) eluting: the composition of the phase that flows in regulation liquid chromatograph, initial eluent consists of the Mobile phase B of 25%, and eluting also collects fraction.After end, the Mobile phase B of regulation eluent composition to 50%, repeated collection process.It is followed successively by the B of the 75% and B of 100% below.
(3) lcms analysis: the fraction collected is concentrated into suitable volume through traditional vacuum concentration systems.And then carry out reversed-phase liquid chromatography mass spectrometry analysis below.
(4) data authentication: the data obtained are analyzed in RPLCMS separation and carries out database search qualification, complete data analysis process.
This technology has been applied in the experiment of overall Separation of Proteins by the present invention, and filler used is the strong cation exchange flavoring agent SCX in ion exchange, achieves good experimental result.In independent one-dimensional reversed phase chromatography is tested, once test our qualification and obtained 21 protein.Applying after said apparatus carries out two dimensional separation, we have obtained 34 protein.It can be seen that the use of our segregation apparatus substantially refer to the quantity of Identification of Fusion Protein.
Claims (2)
1. one kind carries out simple and fast ion exchange fractionating device to polypeptide or protein mixture, including A parts, B parts and C parts, it is characterized in that, described B parts are chromatographic column, are to run through a conduit at two hollow screw centers, and conduit puts in the two-way on left and right both sides, there is female thread at the left and right two of two-way, two hollow screws have external screw thread, are spirally connected with left and right two two-ways respectively, and two hollow screw fronts have sealing ferrum lasso, sieve plate successively；Described A parts are also to run through a conduit at two hollow screw centers, and one of them screw is spirally connected with the left side two-way of B parts, and there is sealing ferrum lasso the screw front of A parts；Described C parts are to be spirally connected with the right two-way in B parts by sealing ferrum lasso with a hollow screw, then collect fraction capillary tube with one, inject in the hollow screw in C parts.
2. the simple and fast ion that carries out polypeptide or protein mixture as described in claim 1 exchanges fractionating device, it is characterised in that the conduit diameter of described A parts is less than the conduit of B parts, and described conduit uses duroplasts to make.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610291376.1A CN106018574A (en) | 2016-05-05 | 2016-05-05 | Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610291376.1A CN106018574A (en) | 2016-05-05 | 2016-05-05 | Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106018574A true CN106018574A (en) | 2016-10-12 |
Family
ID=57081219
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610291376.1A Pending CN106018574A (en) | 2016-05-05 | 2016-05-05 | Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106018574A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6316604B1 (en) * | 1988-04-01 | 2001-11-13 | Avant Immunotherapeutics, Inc. | Human C3b/C4b receptor (CR1) |
| JP2011205964A (en) * | 2010-03-30 | 2011-10-20 | Japan Health Science Foundation | Tendon and/or ligament cell differentiation-inducing agent |
| CN102590402A (en) * | 2012-03-28 | 2012-07-18 | 厦门大学 | Preparing method of capillary tube chromatographic columns |
| CN203329735U (en) * | 2013-04-25 | 2013-12-11 | 齐齐哈尔大学 | Ion exchanging device for minitype laboratory |
| CN105413666A (en) * | 2015-11-24 | 2016-03-23 | 徐州医学院 | Solid-phase extraction filler, solid-phase extraction column and preparation method and application of solid-phase extraction filler |
-
2016
- 2016-05-05 CN CN201610291376.1A patent/CN106018574A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6316604B1 (en) * | 1988-04-01 | 2001-11-13 | Avant Immunotherapeutics, Inc. | Human C3b/C4b receptor (CR1) |
| JP2011205964A (en) * | 2010-03-30 | 2011-10-20 | Japan Health Science Foundation | Tendon and/or ligament cell differentiation-inducing agent |
| CN102590402A (en) * | 2012-03-28 | 2012-07-18 | 厦门大学 | Preparing method of capillary tube chromatographic columns |
| CN203329735U (en) * | 2013-04-25 | 2013-12-11 | 齐齐哈尔大学 | Ion exchanging device for minitype laboratory |
| CN105413666A (en) * | 2015-11-24 | 2016-03-23 | 徐州医学院 | Solid-phase extraction filler, solid-phase extraction column and preparation method and application of solid-phase extraction filler |
Non-Patent Citations (4)
| Title |
|---|
| SHABAZ MOHAMMED等: "Strong cation exchange (SCX) based analytical methods for the targeted analysis of protein post-translational modifications", 《ANALYTICAL BIOTECHNOLOGY》 * |
| 卜春苗等: "强阳离子交换树脂在蛋白质分离纯化中的应用", 《宁夏工程技术》 * |
| 张锴等: "强阳离子交换-加压毛细管点色谱法分离多肽", 《分析试验室》 * |
| 柯从玉: "简易型变性蛋白复性并同时纯化色谱柱的研制及应用", 《中国优秀博硕士学位论文全文数据库 (硕士) 工程科技Ⅰ辑》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6989129B2 (en) | Automated capillary liquid chromatography small volume analysis system | |
| CN103940941B (en) | Automatic pre-treatment-Ultra Performance Liquid Chromatography/mass spectrum on-line analysis system | |
| US9679757B2 (en) | Liquid chromatography systems and methods | |
| US20110247403A1 (en) | Apparatus And Methods For Multidimensional Analysis | |
| Eschelbach et al. | Improved protein recovery in reversed-phase liquid chromatography by the use of ultrahigh pressures | |
| US8074494B2 (en) | Plugged flow 2D chromatography | |
| US20070017868A1 (en) | Fluidic analysis with hydrophobic and hydrophilic compounds trapping | |
| CN105223264A (en) | Mark method, device and application in a kind of simulation of mass spectrum quantitative test | |
| CN109030689B (en) | Capillary chromatographic column pre-assembling preparation method | |
| CN101169391B (en) | Two-dimensional highly effective liquid phase chromatographic system and its uses | |
| CN110208401B (en) | Online analysis system and method of solid phase dehydration extraction-supercritical fluid chromatography-mass spectrometry | |
| CN101206197A (en) | System for on-line desalinization, enrichment and mass spectrum of double split flow noy upgrade liquid chromatogram | |
| Maciel et al. | Towards a universal automated and miniaturized sample preparation approach | |
| PAULAáPACKER et al. | Automated on-line preconcentration for trace metals determination in water samples by inductively coupled plasma mass spectrometry | |
| EP2210087B1 (en) | Method and apparatus for isotope-ratio analysis | |
| CN106018574A (en) | Fractionation device for carrying out simple and rapid ion exchange on peptide or protein mixture | |
| KR101795747B1 (en) | Noncontiguous sample fractionating and concatenating unit and dual online multi-fuctional liquid chronatography device having the same | |
| CN208654099U (en) | Transformer oil gas analysis Special instrument | |
| US20090000358A1 (en) | Method of Analysis Using Chromatographic Pre-Separation | |
| Clarke et al. | The current status and future of two-and multidimensional liquid chromatography in pharmaceutical R&D and QC | |
| van Zelst et al. | Characterising polar compounds using supercritical fluid chromatography–nuclear magnetic resonance spectroscopy (SFC–NMR) | |
| JP2003149218A (en) | Two-dimensional high-performance liquid chromatograph and protein analyzing apparatus using the same | |
| CN111610264A (en) | Hydrophilic interaction chromatography-reversed phase chromatography combined two-dimensional liquid phase system and analysis method thereof | |
| Gaikwad et al. | Ultra performance liquid chromatography: A recent novel development in HPLC | |
| Link et al. | Analysis of protein composition using multidimensional chromatography and mass spectrometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161012 |