CN104344985B - A kind of magnetic separation device - Google Patents
A kind of magnetic separation device Download PDFInfo
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- CN104344985B CN104344985B CN201310347133.1A CN201310347133A CN104344985B CN 104344985 B CN104344985 B CN 104344985B CN 201310347133 A CN201310347133 A CN 201310347133A CN 104344985 B CN104344985 B CN 104344985B
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Abstract
The invention discloses a kind of extraction of order, the magnetic separation device of continuous elution micro substance, described device includes：Disc sample holder, fan-shaped sample disc, sample cell and bar magnet.The magnetic separation device convenience easy to use of the present invention; the magnetisable material that can be used in quick separating tubular container, there is important application value in research fields such as immunocyte separation, protein science separation, immobilized enzyme, heavy metal separation, biological medicine engineering and environmental protection.
Description
Technical field
The invention belongs to separation technology field, is on a kind of extraction of order, continuous elution micro substance specifically
Magnetic separation device.
Background technology
Biomarker (biomarker) refers to a kind of physiology, pathology or the therapeutic process for being available for objective determination and evaluation
In certain characteristic biochemical indicator, can be from Molecular level study pathogenesis, in accurate, sensitively evaluation early stage, low water
There is the advantage of uniqueness in terms of flat infringement, it is possible to provide early warning, largely provide auxiliary diagnosis for clinician
Foundation.DNA, RNA, protein, micromolecule polypeptide and saccharide compound have been widely used in basis as molecular marked compound
Research and medical diagnosis on disease research.The protein-based biomarker that important vital movement is undertaken in life entity is often low abundance
Albumen, but extremely low content carrys out extreme difficulties to subsequent analysis and detection band, limits the research to it and understanding, has biology
The analysis of the low-abundance protein of label function has become the emphasis and difficult point content of protein science research with identification.Therefore egg
The isolation technics of white matter group and peptide group, particularly extraction has biomarker from highly complex and dynamic atmosphere biological sample
The low-abundance phosphorylation of function, modified albumen or the endogenous peptide fragment such as glycosylate, methylate, being proteomics and peptide group
Learn the primary final steps of research.
Fixing metal ions affinity chromatography (IMAC) technology and metal oxide affinity chromatography (MOAC) technology are peculiar because of its
Selective enrichment ability, developed rapidly in modified protein, the enrichment of peptide fragment and separation field, but blood sample, urine sample, lymph, ridge
PH value, ion concentration, amino acid after the complicated substrate sample treatment such as marrow, milk, tissue extract, vegetable seeds and highly concentrated
Degree albumen such as covers at the presence of factor so that the enrichment and separation process of IMAC technologies and MOAC technologies in modified protein, peptide fragment
In be restricted.Research shows there is very big complementarity therebetween, and the factor such as pH of eluent has for elution effect
Significantly affect, such as when being eluted on IMAC, mono-phosphorylated peptide fragment is eluted in pH=1.0, and multi-phosphopeptide
Section needs to elute under alkaline environment.
Fe with superparamagnetism3O4Microballoon, many fields being applied to based on its own characteristic in biomedicine, such as
Magnetic resonance imaging, targeted drug delivery, cell and Separation of Proteins, immune detection and treatment of cancer etc..So far, reported
It is a variety of that magnetic ball surface is modified into different chelation groups or coats different metal oxides, then using IMAC or MOAC
Technical principle is used for the enrichment of modified albumen, immunocyte etc. with separating.
The content of the invention
Present inventor has invented a kind of order that different target material in micro-example is realized based on magnetic material
Extraction, the magnetic separation device of continuous elution micro substance, separator convenience easy to use, can be used for quick separating pipe
Magnetisable material in shape container, in immunocyte separation, protein science separation, immobilized enzyme, heavy metal separation, biological medicine engineering
And the research field such as environmental protection has important application value.
Therefore, it is an object of the invention to provide a kind of extraction of order, the magnetic separation device of continuous elution micro substance.
The present invention is realized by following technical scheme.
A kind of order extraction, the magnetic separation device of continuous elution micro substance, described device include：Disc sample branch
Frame, fan-shaped sample disc, sample cell and bar magnet.
According to the present invention, the fan-shaped sample disc uniformly, stably stationary arrangement in the disc sample holder, and
Can conveniently it remove.
According to the present invention, the fan-shaped sample disc is 6 groups.
According to the present invention, the sample cell is radially arranged centered on the center of the disc sample holder.
According to the present invention, 4 groups of sample cells, any two adjacent fan-shaped samples are uniformly distributed in each fan-shaped sample disc
Angle of radiation between two groups of sample cells of product disk is identical.
According to the present invention, the bar magnet contains sleeve pipe.
According to the present invention, the bottom of described sleeve pipe is cone-shaped.
According to the present invention, the bar magnet is 3 groups.
According to the present invention, in 60 ° and 120 ° arrangements between the bar magnet, fan-shaped sample where different fiber materials is correspond to respectively
The initial position of product disk.
According to the present invention, the disc sample holder and the bar magnet are controlled by stepper motor, realize the fan-shaped sample
360 ° between product disk and the bar magnet rotate against.
Beneficial effect：The order extraction of the present invention that different target material in micro-example is realized based on magnetic material
Take, continuously elute micro substance magnetic separation device especially suitable for immunocyte separation, protein science separation, immobilized enzyme,
Generally there was only 50-500 μ L samples in heavy metal separation, biological medicine engineering, need to identify point of the biochemical analysis of many indexes
From field, sample is avoided repeatedly to shift caused sample loss and cross pollution.In protein science research, it can be used for phosphorus
The complementary enrichment of the equivalent class protein of acidifying modification and peptide fragment is washed with mono-phosphorylated peptide fragment and the continuous of multi-phosphopeptide section
It is de-, it can also be used to phosphorylation, the enrichment and elution variety classes modified protein and the peptide fragment such as glycosylate, methylate, it may also be used for
Segmented enrichment and elution are completed according to the molecular weight distribution of protein.
Brief description of the drawings
Fig. 1 is disc sample holder, fan-shaped sample disc, the layout of sample cell；
Fig. 2 is same plane, the partial schematic diagram of three row's bar magnets of radial arrangement；
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.It should be understood that following examples are merely to illustrate this
Invention is not for restriction the scope of the present invention.
The technical principle of the device of the present invention：By rotating against for circular sample disk and three groups of bar magnets, it is respectively completed micro-
Measure the order extraction of the object in sample and continuous elution.
Embodiment 1, order extraction, continuous washing device
As shown in figure 1, the order extraction of the present invention, continuous washing device are mainly made up of four parts：Disc sample branch
Frame, fan-shaped sample disc, sample cell and three row's bar magnets (containing sleeve pipe).Wherein, disc sample holder (or three row's bar magnets) needs
Stepper motor controls, and realizes that 360 between fan-shaped sample disc and bar magnet ° rotate against.Fan-shaped sample disc is plastic material, Ke Yiwen
Surely it is fixed on disc sample holder, and can conveniently removes.Sample cell need to select polytetrafluoroethylene (PTFE) etc. will not cause cell,
The hard material of the denaturation such as protein, biology enzyme, sample cell internal diameter are about 6-10mm, and depth is about 5-8cm, and can be inserted with seamless
Enter in fan-shaped sample disc, circular sample support, fan-shaped sample disc and sample cell three closely, stably will can be nested in one
Rise, it is impossible to rock.
As shown in Fig. 2 the arrangement position of three row's bar magnets is in 60 ° and 120 ° of radial arrangements respectively, enrichment material is corresponded to respectively
The initial position of material.
As shown in figure 3, the external diameter of bar magnet (containing sleeve pipe) should 2mm thin compared with sample cell internal diameter, Mechanical course bar magnet (contains sleeve pipe)
Sample cell depth capacity is inserted to be advisable apart from sample cell bottom 1-2mm；Magnetic rod sleeve material requires identical with sample cell material,
Thickness is no more than 0.3mm, and bottom is centrum shape, pyramidal portion height 1-2mm, bar magnet Surface field>1500 Gausses, bar magnet requirement
Magnetic is uniform, and shape is identical with casing inner diameter, easily can be moved up and down in sleeve pipe, and gap therebetween should be less than 0.5mm,
Mechanical course bar magnet can be fully removed sleeve pipe, or bar magnet and sleeve pipe synchronously remove sample cell together.In use, bar magnet is put
In 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, transfer flow of the sorbing material between different sample cells is shown in Fig. 4, sorbing material is placed in dress first
Have in the sample cell of sample and be enriched with target substance, before being then transferred into after being adsorbed in the sample cell equipped with cleaning fluid, extracting
Cleaning, be finally transferred to that sample elution gets off in the elution bottle equipped with eluent.
Embodiment 2, application
The Magnetic Isolation method that three kinds of enrichment material (a, b, c) order extractions, each personal two kinds of eluents continuously elute is such as
Under.
2.1st, 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 extraction；
B2、C2：Sample buffer or deionized water；
EA0：Cleaning solution corresponding to a enrichment materials, for the washing after extraction, before elution；
EA1、EA2：Two kinds of eluents corresponding to a enrichment materials, for continuously eluting；
EB0：Cleaning solution corresponding to b enrichment materials, for the washing after extraction, before elution；
EB1、EB2：Two kinds of eluents corresponding to b enrichment materials, for continuously eluting；
EC0：Cleaning solution corresponding to c enrichment materials, for the washing after extraction, before elution；
EC1、EC2：Two kinds of eluents corresponding to c enrichment materials, for continuously eluting；
Liquid level is no more than the 75% of sample cell height after applied sample amount requires the fully-inserted sample cell of magnetic rod sleeve.
2.2nd, order extraction, continuous elution
2.2.1, a enrichment materials extract, and b enrichment materials prepare
After controlling a enrichment materials to sequentially pass through sample-loading buffer A2 activation by stepper motor, into sample position S, through magnetic
Rod sleeve pipe shakes repeatedly, after the completion of selective enrichment, bar magnet insertion, a enrichment materials of selective absorption target substance is adsorbed
To sleeve bottom centrum position.Now b enrichment materials prepare into B area.
2.2.2, a enrichment materials elute, and b enrichment materials extraction, c enrichment materials prepare
Stepper motor controls bar magnet and circular sample disk to relatively rotate, and enters a enrichment materials for being adsorbed with target substance
EA regions, synchronous b enrichment materials enter the operation of a-quadrant, repeatedly 2.2.1, the target substance that a enrichment materials adsorb successively is existed
Two EA0 positions are completed to clean twice, and elution is successively sequentially then completed in EA1 and EA2 according to the different of property；B is rich simultaneously
Collect material to complete to be directed to the extraction of target substance in sample.Now c enrichment materials prepare into C regions.
2.2.3, b enrichment materials elute, the extraction of c enrichment materials
Stepper motor controls bar magnet and circular sample disk to relatively rotate, and enters the b enrichment materials for being adsorbed with target substance
EB regions, synchronous c enrichment materials enter the operation of a-quadrant, repeatedly 2.2.1, the target substance that b enrichment materials adsorb successively is existed
Two EB0 positions are completed to clean twice, and elution is successively sequentially then completed in EB1 and EB2 according to the different of property；C is rich simultaneously
Collect material to complete to be directed to the extraction of target substance in sample.
2.2.4, c enrichment materials elute
Stepper motor controls bar magnet and circular sample disk to relatively rotate, and enters the c enrichment materials for being adsorbed with target substance
The operation in EC regions, repeatedly 2.2.1, the target substance that c enrichment materials adsorb is set successively to complete to clean twice two EC0 positions,
Then elution is successively sequentially completed in EC1 and EC2 according to the different of property；A enrichment materials and b enrichment materials return respectively simultaneously
To EA regions and a-quadrant, with the simultaneously operating of bar magnet, the elution result in EA regions is not interfered with.
So far, it is fully completed by the order extraction of three kinds of materials, continuous elution process.
If only with two kinds of extractions of extractant order, continuous elutions, will need to be only put into two fan-shaped sample discs of C, EC
Deionized water or sample buffer.
Claims (8)
1. a kind of magnetic separation device, it is characterised in that described device includes：Disc sample holder, fan-shaped sample disc, sample
Pond and bar magnet；Uniformly, stably stationary arrangement in the disc sample holder, and can conveniently take the fan-shaped sample disc
Under；The sample cell is radially arranged centered on the center of the disc sample holder；The bar magnet inserts the sample
In product pond.
2. magnetic separation device according to claim 1, it is characterised in that the fan-shaped sample disc is 6 groups.
3. magnetic separation device according to claim 1, it is characterised in that be uniformly distributed in each fan-shaped sample disc
4 groups of sample cells, the angle of radiation between two groups of sample cells of any two adjacent fan-shaped sample discs are identical.
4. magnetic separation device according to claim 1, it is characterised in that the bar magnet contains sleeve pipe.
5. magnetic separation device according to claim 4, it is characterised in that the bottom of described sleeve pipe is cone-shaped.
6. magnetic separation device according to claim 4, it is characterised in that the bar magnet is 3 groups.
7. magnetic separation device according to claim 4, it is characterised in that in 60 ° and 120 ° arrangements between the bar magnet, divide
The initial position of fan-shaped sample disc where not correspond to different fiber materials.
8. magnetic separation device according to claim 1, it is characterised in that the disc sample holder and the bar magnet
Controlled by stepper motor, realize that 360 ° between the fan-shaped sample disc and the bar magnet rotate against.
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| CN201310347133.1A CN104344985B (en) | 2013-08-09 | 2013-08-09 | A kind of magnetic separation device |
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| CN201310347133.1A CN104344985B (en) | 2013-08-09 | 2013-08-09 | A kind of magnetic separation device |
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| CN104344985B true CN104344985B (en) | 2018-04-03 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105527150B (en) * | 2015-12-10 | 2018-10-23 | 南京医科大学 | A kind of high-throughput magnetic extracting and enriching device and enrichment method |
| CN105543090B (en) * | 2016-01-19 | 2018-08-10 | 山西振东基赛生物科技有限公司 | A kind of dehydration device and method |
| CN110327659B (en) * | 2019-07-19 | 2021-09-24 | 苏州艾捷博雅生物电子科技有限公司 | Extraction and purification method and device based on magnetic extraction material |
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