CN104344985A - Magnetic separation device capable of sequentially extracting and continuously eluting trace substances - Google Patents
Magnetic separation device capable of sequentially extracting and continuously eluting trace substances Download PDFInfo
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- CN104344985A CN104344985A CN201310347133.1A CN201310347133A CN104344985A CN 104344985 A CN104344985 A CN 104344985A CN 201310347133 A CN201310347133 A CN 201310347133A CN 104344985 A CN104344985 A CN 104344985A
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Abstract
The invention discloses a magnetic separation device capable of sequentially extracting and continuously eluting trace substances. The device comprises a disc-shaped sample bracket, a fan-shaped sample plate, a sample tank and a magnetic bar. The magnetic separation device is quick and convenient to use, is capable of quickly separating the magnetic substances in a tubular container, and has important application values in research fields in immune cell separation, proteomics separation, immobilized enzymes, separation of heavy metals, biomedical engineering and environmental protection.
Description
Technical field
The invention belongs to separation technology field, specifically, is the magnetic separation device about a kind of order extraction, continuously wash-out micro substance.
Background technology
Biomarker (biomarker) refers to certain the distinctive biochemical indicator in a kind of physiology, pathology or the therapeutic process that can supply objective determination and evaluation, can from Molecular level study pathogenesis, evaluating the advantage having uniqueness in early stage, low-level infringement accurately, sensitively, early warning can be provided, to a great extent for clinician provides the foundation of auxiliary diagnosis.DNA, RNA, protein, micromolecule polypeptide and saccharide compound have been widely used in fundamental research and medical diagnosis on disease research as molecular marked compound.The protein-based biomarker low-abundance protein often of important vital movement is born in life entity, but extremely low content brings extreme difficulties to subsequent analysis and detection, limit the research to it and understanding, the identification with the low-abundance protein of biomarker function has become the Focal point and difficult point content of protein science research.Therefore the isolation technics of Leaf proteins and peptide group, particularly extract from high complexity and dynamic atmosphere biological sample there is biomarker function low-abundance phosphorylation, glycosylation, modified albumen or the endogenous peptide section such as methylate, and is the primary final steps that proteomics and peptide group are studied.
Fixing metal ions affinity chromatography (IMAC) technology and metal oxide affinity chromatography (MOAC) technology are because of its distinctive selective enrichment ability, in enrichment and the separation field develop rapidly of modified protein, peptide section, but the pH value after the complicated substrate sample preparation such as blood sample, urine sample, lymph, spinal cord, milk, tissue extract, vegetable seeds, ion concentration, amino acid and high concentration albumen such as to cover at the existence of factor, make IMAC technology and MOAC technology be restricted in the enrichment and detachment process of modified protein, peptide section.Research shows there is very large complementarity therebetween, and the factor such as the pH of eluent has a significant effect for elute effect, such as on IMAC during wash-out, mono-phosphorylated peptide section is all eluted when pH=1.0, and multi-phosphopeptide section needs wash-out under alkaline environment.
There is the Fe of superparamagnetism
3o
4microballoon, has been applied to the many fields in biomedicine based on himself characteristic, as magnetic resonance imaging, targeted drug delivery, cell and Separation of Proteins, immune detection and treatment of cancer etc.Up to now, reported multiple by chelation groups different for the finishing of magnetic ball or coated different metal oxide, the enrichment then adopting IMAC or MOAC know-why to be used for modified albumen, immunocyte etc. be separated.
Summary of the invention
Present inventor has invented a kind of magnetic separation device realizing order extraction, continuously the wash-out micro substance of different target material in micro-example based on magnetic material; this tripping device convenience easy to use; may be used for the magnetisable material in quick separating tubular container, in research fields such as immunocyte separation, protein science separation, immobilized enzyme, heavy metal separation, biological medicine engineering and environmental protection, there is important using value.
Therefore, the object of the present invention is to provide the magnetic separation device of the extraction of a kind of order, continuously wash-out micro substance.
The present invention is realized by following technical scheme.
A magnetic separation device for order extraction, continuously wash-out micro substance, described device comprises: disc sample holder, fan-shaped sample disc, sample cell and bar magnet.
According to the present invention, described fan-shaped sample disc evenly, stably stationary arrangement in described disc sample holder, and can conveniently to take off.
According to the present invention, described fan-shaped sample disc is 6 groups.
According to the present invention, described sample cell, centered by the center of described disc sample holder, arranges radially.
According to the present invention, each described fan-shaped sample disc is uniformly distributed 4 groups of sample cells, the angle of radiation between two groups of sample cells of any two adjacent fan-shaped sample disc is identical.
According to the present invention, described bar magnet is containing sleeve pipe.
According to the present invention, the bottom of described sleeve pipe is cone-shaped.
According to the present invention, described bar magnet is 3 groups.
According to the present invention, in 60 ° and 120 ° of arrangements between described bar magnet, correspond to the initial position of the fan-shaped sample disc in different fiber material places respectively.
According to the present invention, described disc sample holder and described bar magnet are controlled by step motor, realize the rotation relative to 360 ° between described bar magnet of described fan-shaped sample disc.
Beneficial effect: order extraction, the continuously magnetic separation device of wash-out micro substance realizing different target material in micro-example based on magnetic material of the present invention is specially adapted to immunocyte separations, protein science separations, immobilized enzyme, heavy metal are separated, the separation field that usually only has 50-500 μ L sample in biological medicine engineering, need the biochemical analysis of qualification many indexes, avoids sample repeatedly to shift caused sample loss and cross pollution.In protein science research, may be used for phosphorylation modification and be equal to the complementary enrichment of class protein and peptide section and the continuous wash-out of mono-phosphorylated peptide section and multi-phosphopeptide section, also can be used for phosphorylation, glycosylation, the enrichment of variety classes modified protein and the peptide section such as to methylate and wash-out, also can be used for completing sectional type enrichment and wash-out according to the molecular weight distribution of protein.
Accompanying drawing explanation
Fig. 1 is the layout of disc sample holder, fan-shaped sample disc, sample cell;
The partial schematic diagram that Fig. 2 is same plane, radiate three arranges bar magnet;
Fig. 3 is the schematic diagram of sample cell, bar magnet and sleeve pipe;
Fig. 4 is the schematic diagram of transfer magnetite gathering material movement;
Fig. 5 is the schematic diagram of initial loading position.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described.Should be understood that following examples only for illustration of the present invention but not for limiting scope of the present invention.
The know-why of device of the present invention: by the relative rotation of circular sample dish with three groups of bar magnets, completes the order extraction of the object in micro-example and continuous wash-out respectively.
Embodiment 1, order extraction, continuously washing device
As shown in Figure 1, order extraction, continuously washing device of the present invention form primarily of four parts: disc sample holder, fan-shaped sample disc, sample cell and three rows' bar magnet (containing sleeve pipe).Wherein, disc sample holder (or three row's bar magnets) needs step motor to control, and realizes fan-shaped sample disc and ° relative rotation of 360 between bar magnet.Fan-shaped sample disc is plastic material, stably can be fixed on disc sample holder, and can conveniently take off.Sample cell need select teflon etc. can not cause the hard material of the sex change such as cell, protein, biology enzyme, sample cell internal diameter is about 6-10mm, the degree of depth is about 5-8cm, and can in the fan-shaped sample disc of seamless insertion, circular sample support, fan-shaped sample disc and sample cell three want can be tight, stable nest together, can not rock.
As shown in Figure 2, the arrangement position of three row's bar magnets is 60 ° and 120 ° of radiate, the respectively initial position of corresponding enrichment material respectively.
As shown in Figure 3, the external diameter of bar magnet (containing sleeve pipe) should the comparatively thin 2mm of sample cell internal diameter, and Mechanical course bar magnet (containing sleeve pipe) inserts sample cell depth capacity and is advisable with 1-2mm bottom distance sample cell; Magnetic rod sleeve material requires identical with sample cell material, thickness is no more than 0.3mm, bottom is centrum shape, pyramidal portion height 1-2mm, bar magnet Surface field >1500 Gauss, bar magnet requires that magnetic is even, shape is identical with casing inner diameter, and can easily move up and down in sleeve pipe, gap therebetween should be less than 0.5mm, Mechanical course bar magnet can shift out sleeve pipe completely, or bar magnet synchronously shifts out sample cell together with sleeve pipe.During use, be placed in by bar magnet in sleeve pipe, sleeve pipe is placed in sample cell, and enrichment material is adsorbed in the cone position of sleeve bottom by bar magnet.
As shown in Figure 4, the transfer flow of sorbing material between different sample cell is shown in Fig. 4, first sorbing material is placed in the sample cell enrich target material that sample is housed, then transfer in the sample cell that cleaning fluid is housed the cleaning after carrying out absorption, before extraction, finally transfer in the wash-out bottle that eluent is housed sample elution to get off.
Embodiment 2, application
(the magnetic resolution method of a, b, c) order extraction, each personal two kinds of continuous wash-outs of eluent is as follows for three kinds of enrichment materials.
2.1, loading
As shown in Figure 5, each sample pond is respectively according to following requirements loading.
S: sample position, for placing primary sample;
A1, B1, C1: enrichment material a, b, c;
A2: sample buffer, for the washing before extracting;
B2, C2: sample buffer or deionized water;
The cleansing solution that EA0:a enrichment material is corresponding, for the washing after extracting, before wash-out;
Two kinds of eluents that EA1, EA2:a enrichment material is corresponding, for continuous wash-out;
The cleansing solution that EB0:b enrichment material is corresponding, for the washing after extracting, before wash-out;
Two kinds of eluents that EB1, EB2:b enrichment material is corresponding, for continuous wash-out;
The cleansing solution that EC0:c enrichment material is corresponding, for the washing after extracting, before wash-out;
Two kinds of eluents that EC1, EC2:c enrichment material is corresponding, for continuous wash-out;
After applied sample amount requires magnetic rod sleeve to insert sample cell completely, liquid level is no more than 75% of sample cell height.
2.2, order extraction, continuously wash-out
2.2.1, a enrichment material extraction, b enrichment material prepare
After sequentially passing through sample-loading buffer A2 activation by step motor control a enrichment material, enter sample position S, repeatedly shake through magnetic rod sleeve, after selective enrichment completes, bar magnet inserts, and a enrichment material of selective adsorption target substance is adsorbed to sleeve bottom centrum position.Now b enrichment material enters the preparation of B region.
2.2.2, a enrichment material wash-out, b enrichment material extract, c enrichment material prepare
Step motor controls bar magnet and circular sample dish relatively rotates, the a enrichment material being adsorbed with target substance is made to enter EA region, synchronous b enrichment material enters a-quadrant, the operation of 2.2.1 repeatedly, the target substance that a enrichment material is adsorbed successively completes twice cleaning two EA0 positions, then successively in EA1 and EA2, sequentially completes wash-out according to the different of character; B enrichment material completes the extraction for target substance in sample simultaneously.Now c enrichment material enters the preparation of EC region.
2.2.3, b enrichment material wash-out, c enrichment material extract
Step motor controls bar magnet and circular sample dish relatively rotates, the b enrichment material being adsorbed with target substance is made to enter EB region, synchronous c enrichment material enters a-quadrant, the operation of 2.2.1 repeatedly, the target substance that b enrichment material is adsorbed successively completes twice cleaning two EB0 positions, then successively in EB1 and EB2, sequentially completes wash-out according to the different of character; C enrichment material completes the extraction for target substance in sample simultaneously.
2.2.4, c enrichment material wash-out
Step motor controls bar magnet and circular sample dish relatively rotates, the c enrichment material being adsorbed with target substance is made to enter EC region, the operation of 2.2.1 repeatedly, the target substance that c enrichment material is adsorbed successively completes twice cleaning two EC0 positions, then successively in EC1 and EC2, sequentially completes wash-out according to the different of character; A enrichment material and b enrichment material get back to EA region and a-quadrant respectively simultaneously, along with the synchronous operation of bar magnet, can not affect the wash-out result in EA region.
So far, the order through three kinds of materials extracts, continuous elution process all completes.
If only adopt the extraction of two kinds of extractant orders, continuously wash-out, only deionized water or sample buffer need be put into by C, EC two fan-shaped sample disc.
Claims (10)
1. a magnetic separation device for order extraction, continuously wash-out micro substance, it is characterized in that, described device comprises: disc sample holder, fan-shaped sample disc, sample cell and bar magnet.
2. magnetic separation device according to claim 1, is characterized in that, described fan-shaped sample disc evenly, stably stationary arrangement in described disc sample holder, and can conveniently to take off.
3. magnetic separation device according to claim 2, is characterized in that, described fan-shaped sample disc is 6 groups.
4. magnetic separation device according to claim 1, is characterized in that, described sample cell, centered by the center of described disc sample holder, arranges radially.
5. magnetic separation device according to claim 4, is characterized in that, each described fan-shaped sample disc is uniformly distributed 4 groups of sample cells, and the angle of radiation between two groups of sample cells of any two adjacent fan-shaped sample disc is identical.
6. magnetic separation device according to claim 1, is characterized in that, described bar magnet is containing sleeve pipe.
7. magnetic separation device according to claim 1, is characterized in that, the bottom of described sleeve pipe is cone-shaped.
8. magnetic separation device according to claim 6, is characterized in that, described bar magnet is 3 groups.
9. magnetic separation device according to claim 6, is characterized in that, in 60 ° and 120 ° of arrangements between described bar magnet, correspond to the initial position of the fan-shaped sample disc in different fiber material places respectively.
10. magnetic separation device according to claim 1, is characterized in that, described disc sample holder and described bar magnet are controlled by step motor, realizes the rotation relative to 360 ° between described bar magnet of described fan-shaped sample disc.
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Cited By (3)
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| CN105527150A (en) * | 2015-12-10 | 2016-04-27 | 南京医科大学 | High-throughput magnetic extraction enrichment device and enrichment method |
| CN105543090A (en) * | 2016-01-19 | 2016-05-04 | 山西振东基赛生物科技有限公司 | Dehydration device and method |
| CN110327659A (en) * | 2019-07-19 | 2019-10-15 | 天津迪沃特生物电子科技有限公司 | A kind of extraction and purification method and device based on magnetic fiber material |
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| CN104344985B (en) | 2018-04-03 |
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