KR20160133826A - Apparatus for separating fine endoplasmic reticulum using electrophoresis - Google Patents

Apparatus for separating fine endoplasmic reticulum using electrophoresis Download PDF

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KR20160133826A
KR20160133826A KR1020150066864A KR20150066864A KR20160133826A KR 20160133826 A KR20160133826 A KR 20160133826A KR 1020150066864 A KR1020150066864 A KR 1020150066864A KR 20150066864 A KR20150066864 A KR 20150066864A KR 20160133826 A KR20160133826 A KR 20160133826A
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flow channel
biological sample
electrolyte
electrophoresis
supply unit
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KR1020150066864A
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Korean (ko)
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천홍구
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고려대학교 산학협력단
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Publication of KR20160133826A publication Critical patent/KR20160133826A/en

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
    • B01L9/527—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for microfluidic devices, e.g. used for lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0421—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic electrophoretic flow

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The present invention relates to a device for separating fine endoplasmic reticulum by using electrophoresis and, more specifically, to a device for separating fine endoplasmic reticulum by using electrophoresis, in which a biological sample is separated according to electrophoresis fluidity. To this end, the device of the present invention comprises: a fluidic channel in which a biological sample and a buffer separately and horizontally flow along one direction; a first feeding port connected with a first route in one end of the fluidic channel, and feeding the buffer therein; a second feeding port connected with a second route in one end of the fluidic channel, and feeding the biological sample therein; an electrical field generating unit for generating an electrical field in a direction vertically crossing the one direction in the fluidic channel to separate the biological sample flowing along the fluidic channel according to electrophoresis fluidity; and a plurality of discharging ports connected with the other end of the fluidic channel to separately flow samples separated according to electrophoresis fluidity. A large amount of samples are continuously separated according to electrophoresis fluidity, so damage to fine endoplasmic reticulum by a biological sample can be prevented.

Description

[0001] Apparatus for separating fine endoplasmic reticulum using electrophoresis [

The present invention relates to a microfibrillar separator, and more particularly, to a microfibrillar separator using electrophoresis for separating a biological sample according to electrophoretic fluidity.

In recent years, interest and research on biotechnology have been actively pursued. However, existing bioanalytical systems are difficult to rapidly process rapidly growing bio information. Therefore, the biological detection system for the identification of life phenomena and drug development and diagnosis is based on microfluidics, and a micro-comprehensive analysis system (μ-TAS : micro-Total Analysis System) and lab-on-a-chip. Since most of the biochemical samples to be analyzed are present in solution, the technique of delivering liquid samples is the most important factor. Microfluidics is a research field for controlling the flow of such microfluidics, and is a field for research and development of core technologies that are based on commercialization of the microcomputer analysis system and lab-on-a-chip.

The micro total analysis system is a system that comprehensively implements chemical and biological experiments and analyzes, which are subjected to a plurality of experimental steps and reactions, on one unit existing on one laboratory. Such a micro total analysis system is composed of a sampling region, a microfluidic circuit, a detector, and a controller for controlling them.

Also, the lab-on-a-chip means that the concept and function of the micro-comprehensive analysis system are implemented on a single chip in the meaning of a 'laboratory on a chip'. Therefore, in order to develop the lab-on-a-chip, a circuit is formed with microchannels through which a solution can flow on the surface of plastic, glass, or silicon, and then pretreatment, separation, dilution, mixing, biochemical reaction, Chip and integrated on a chip of a semiconductor device.

On the other hand, in vivo micro-vesicles (micro vesicles) are small vesicles of membrane structure that are present in various kinds of cells or secreted from cells. Microvesicles secreted out of the cell are (1) exosomes: membrane vesicles of 30-100 nm in diameter originated from the origin of the bacteria, (2) shedding microvesicles (SMVs): flowing directly from the plasma membrane (3) Apoptotic blebs: vesicles having a diameter of 50 to 5000 nm, which are discharged by dying cells.

The in vivo micro-vesicles (microvesicles), such as exosomes, are vesicles of the size of a few tens of nanometers secreted from the cells, and are produced in the cytoplasm or cells inside the lipid bilayer or lipid monolayer It is a structure containing protein and RNA. Exosomes are a means of intercellular communication through the exchange of proteins and RNAs. They also function to release unnecessary substances in cells. They contain microRNAs (microRNAs and miRNAs) It can be used as a useful marker in diagnosis. Although the importance and the value of the in vivo micro-endoplasmic reticulum as described above have been revealed, it is difficult to obtain the micro-endoplasmic reticulum.

The method of isolating the existing microbejicle is a method of immune-capturing and isolating the microbequicle by combining the microbezyme and the antibody. Such a method may cause a bias depending on the separation or detection target due to masking of antibody recognition sites due to changes in the protein structure, microbial heterogeneity, protein interaction, and the like. Complex processes or expensive equipment may be required for separation or detection, and sample consumption may be high. Therefore, it is necessary to efficiently separate microbeads from a small amount of sample, independent of the target.

In addition, in order to separate the microbejicles or exosomes, the centrifugal separation method was generally used. A solution of Ficoll solution or OptiPrep (Nycomed Pharma, Norway) or the like was added to the cell or tissue sample solution and centrifuged to obtain microbicule. However, this method requires a large amount (volume) of sample as well as requiring pretreatment of the cell or tissue sample liquid. Requires several centrifugation processes, requires special reagents and devices for centrifugation, and takes a lot of time and costs. As a result, the pellet containing the microbeques obtained through the centrifugation contains a lot of impurities such as fine protein molecules and cell debris similar in density and mass to the microbequicle. In addition, since the impurities do not differ greatly in density from the microvacles, they are not easily separated and thus are not applicable to on-site diagnostics that require immediate response. In addition, microvessels to be obtained may be damaged due to high inertial force for a long time, which may cause fundamental problems in studying biological reactions.

Therefore, it is necessary to develop a new system capable of continuously separating micro vesicles without damaging micro vesicles from biological samples.

Korean Patent Publication No. 2005-0119128

SUMMARY OF THE INVENTION The present invention has been conceived to solve the above-mentioned problems, and an object of the present invention is to provide an electrophoretic mobility electrophoretic mobility electrophoretic mobility analyzer capable of selectively obtaining microbubbles of a desired size without damaging microbubbles from a biological sample. And to provide a device for separating a microfibrillar body using the same.

To this end, the apparatus for separating micro-fibrils using electrophoresis according to the present invention comprises a flow channel in which a biological sample and a buffer are separated and flow side by side along one direction; A first injection port connected to a first path at one end of the flow channel to inject the buffer; A second injection port connected to a second path at one end of the flow channel to inject the biological sample; An electric field forming unit for forming an electric field in the flow channel in a direction orthogonal to the one direction so that the biological sample flowing along the flow channel is separated according to electrophoretic fluidity; And a plurality of discharge portions connected to the other end of the flow channel such that the separated samples are flowed from each other in accordance with the electrophoretic fluidity, wherein the biological sample is a protein, a microfibril or a mixture thereof.

And the flow channel according to various embodiments of the present invention adjusts the flow rate of the buffer to be injected such that the biological sample flows side by side along the one side wall of the flow channel separately from the buffer.

Also, the flow rate of the injected buffer according to various embodiments of the present invention is 1 to 20 times the flow rate of the biological sample injected.

Meanwhile, the electric field forming unit according to various embodiments of the present invention includes: a cation supply unit connected to one side of the flow channel and supplying cations to the flow channel; An anion supply unit connected to the flow channel so as to be disposed on the opposite side of the cation supply unit via the flow channel, the anion supply unit supplying an anion to the flow channel; And a power supply unit having a pair of electrodes, one for each of the positive ion supply unit and the negative ion supply unit; .

In addition, the electric field forming unit according to various embodiments of the present invention allows particles contained in the biological sample passing through to be positively charged by the positive ions supplied from the positive ion supplying unit, The fibril particles are separated by the electrophoretic fluidity while flowing by an electric field formed in a direction orthogonal to the one direction.

In addition, the cation supply unit according to various embodiments of the present invention may include a first storage unit in which the first electrolyte is stored; And a negative charge polymer selectively transmitting only positive ions among the ions contained in the first electrolyte; Wherein the anion supply unit includes: a second reservoir storing a second electrolyte; A positive charge polymer that selectively transmits only anions among the ions contained in the second electrolyte; .

Meanwhile, the first electrolyte according to various embodiments of the present invention includes a hydrogen ion (H + ), and the second electrolyte includes a hydroxide ion (OH - ).

And the first electrolyte according to various embodiments of the present invention is HCl and the second electrolyte is KOH.

Prior to that, terms and words used in the present specification and claims should not be construed in a conventional and dictionary sense, and the inventor may properly define the concept of the term in order to best explain its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

The apparatus for separating micro-fibrils using electrophoresis according to various embodiments of the present invention has an effect of preventing damage to micro vesicles from a biological sample by continuously separating a large amount of samples according to electrophoresis fluidity.

1 is a conceptual diagram showing a principle of a microfibrillator separation apparatus according to the present invention.
FIG. 2 is a schematic configuration diagram of a microfibrillator separation apparatus according to an embodiment of the present invention; FIG.
3 is a schematic configuration diagram of a microfibril separation apparatus according to another embodiment of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The objectives, specific advantages and novel features of the present invention will become more apparent from the following detailed description taken in conjunction with the accompanying drawings, in which: FIG. It should be noted that, in the present specification, the reference numerals are added to the constituent elements of the drawings, and the same constituent elements are assigned the same number as much as possible even if they are displayed on different drawings. Also, the terms "first "," second ", and the like are used to distinguish one element from another element, and the element is not limited thereto. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In the following description of the present invention, detailed description of related arts which may unnecessarily obscure the gist of the present invention will be omitted.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a conceptual view showing the principle of a microfibrillator separation apparatus according to the present invention.

Referring to FIG. 1, the present invention can separate micro-vesicles or proteins according to electrophoretic mobility using electrophoresis.

On the other hand, the protein or the micro-endoplasmic reticulum particle contained in the biological sample is charged according to the pH. Specifically, the protein or the micro-endoplasmic reticulum particle is positively charged at pH lower than the pI (Isoelectric point) at which the charge becomes zero, do.

In addition, electrophoresis is a phenomenon in which charged particles move in an area where an electric field is applied. The charged particles differ in their flow rates due to factors such as particle size, charge amount, and buffer composition. (electricphoretic mobility). The present invention has been accomplished by using the above-described characteristic, that is, the characteristic that charged particles are separated and flow according to the electrophoretic fluidity.

Herein, the term 'micro-vesicle' refers to a small vesicle of membrane structure existing in various kinds of cells or secreted from cells, and includes extracellular vesicles. The micro-vesicles secreted extracellularly are composed of (1) exosomes: membrane vesicles of 30-100 nm in diameter from the origin of the bacteria, (2) shedding microvesicles (SMVs) (3) Apoptotic blebs: including, but not limited to, vesicles 50-5000 nm in diameter, which are drained by dying cells. The microfilament to be obtained by the present invention may be preferably an exosome.

The 'exosome' is a small vesicle of membrane structure secreted from various kinds of cells. The diameter of the exosome can be as large as 30-1000 nm. Exosomes originate from specific compartments within the cell called multivesicular bodies (MVBs) and are released and secreted out of the cell, rather than being removed directly from the plasma membrane in electron microscopic studies. That is, when fusion of the polycation and the plasma membrane occurs, such vesicles are released into the extracellular environment, which is called exosomes. It is unclear how these exosomes are produced by molecular mechanisms, but it is possible that not only red blood cells but also various types of immune cells and tumor cells, including B-lymphocytes, T-lymphocytes, dendritic cells, platelets and macrophages, It is known to produce and secrete exosome in the state of being. Exosomes are known to be released from many different cell types under normal, pathological, or both conditions.

Biological samples contain protein, particles of similar size to the micro-endoplasmic reticulum, and vesicles of various sizes including micro-endoplasmic reticulum. As the follicle, especially the micro-endoplasmic reticulum, differs in the cell-producing region, the lipid layer constituting the follicle varies in monolayer or bilayer, and the size of the follicle varies.

The biological sample containing the protein and the microfibrillar of the present invention refers to a biological sample from which a desired type of microfibrillar body can be obtained and includes, for example, a body fluid or a cell culture. The body fluid may be at least one selected from the group consisting of urine, mucus, saliva, tears, plasma, serum, urine, sputum, spinal fluid, pleural fluid, aspiration nipple, lymphatic fluid, airway fluid, intestinal fluid, urinary reproductive fluid, , Ascites, cystic tumor body fluids, positive sap or combinations thereof. The cell culture medium means a culture medium from which cells have been removed after cell culture. The composition of the medium may be optionally changed by a person skilled in the art so as to secrete a large amount of micro-endoplasmic reticulum from the cells, but it is preferably a conditioned medium (serum-free medium) or serum .

In addition, the filtration and concentration processes may be arbitrarily added to the sample according to the experimental efficiency desired by the ordinary skilled in the art. The filtration process may be performed by a known filtration method. For example, centrifugation or filtration using a microfilter may be used. The concentration process may be performed by a known concentration process, but is not limited thereto. For example, the process can be performed using a centrifugation method.

The biological sample containing the protein and the microfibrillar of the present invention may preferably be a culture medium after the cell culture or a serum concentrate.

Next, referring to FIGS. 2 and 3, the apparatus 100 for separating a microfibrillar body according to various embodiments of the present invention will be described below.

FIG. 2 is a schematic configuration diagram of a microfibrillator separation apparatus according to an embodiment of the present invention, and FIG. 3 is a schematic configuration diagram of a microfibrillate separation apparatus according to another embodiment of the present invention.

Referring to FIG. 2, the apparatus 110 for separating microfibrillatedglass according to an exemplary embodiment of the present invention includes a flow channel 110 in which a biological sample and a buffer are separated and flow side by side along one direction, A second injection port 130 connected to a second path of the flow channel 110 to inject the biological sample and a second injection port 130 connected to the flow channel 110, An electric field forming unit 140 for forming an electric field in the flow channel 110 in a direction perpendicular to the one direction so that the biological sample flowing along the flow channel 110 is separated according to the electrophoretic fluidity, And a plurality of outlets (150) connected to the other end of the flow channel so that the samples flow from each other.

Preferably, the biological sample is a protein, a micro-vesicle or a mixture thereof.

The flow channel 110 is formed in a buffer, a place where biological samples to be separated are separated and flows side by side, and is formed long in one direction. At this time, it is preferable that the buffer (buffer) flows through the channel and carries and separates the micro-vesicles in the sample, so that it can simultaneously perform a role as a carrier buffer and a partitioning fluid.

The buffer is injected through a first inlet 120 connected to a first path of the flow channel 110 and the biological sample is injected through a second inlet 130 connected to a second path of the flow channel 110, Lt; / RTI > The buffer and the biological sample injected through the injection ports flow along one direction of the flow channel and flow in one direction. As described above, the buffer serves as a carrier buffer and a separation liquid, And flows in parallel. At this time, the biological sample may flow along one side wall of the flow channel 110 to increase the separation effect. The flow can be controlled through adjustment of the flow rate of the buffer injected through the first injection port 120. At this time, the flow rate of the injected buffer is preferably 1 to 20 times the flow rate of the biological sample to be injected. If the upper limit is exceeded, there is a problem that the flow along the one side wall of the sample is unstable. If the flow rate is less than the lower limit value, the sample flows into the discharge outlet 150 before separation There is a problem.

The buffer is not limited in its kind as long as it does not affect the lipid membrane structure of the micro-vesicles contained in the biological sample, which is known in the art. For example, a phosphate buffered saline (PBS), a PBS solution containing sucrose, a PBS solution containing glycine, etc. may be used, and preferably, the solution may be a PBS solution containing sucrose. It is not.

Buffers and biological specimens joining one end of the flow channel 110 and flowing along one direction are separated according to the electrophoretic fluidity while passing through the electric field forming unit 140. The electric field forming unit 140 forms a pH gradient in the flow channel 110 in a direction orthogonal to the one direction so that the electric field forming unit 140 can generate the pH of the protein Or microembossed particles are positively charged. This is because the particles are positively charged at a pH lower than the pH value as described above. Since the region where the particles flow along one side wall of the flow channel corresponds to the acid region having a very low pH, So that a positive charge is generated.

In addition, the electric field forming unit 140 has an electric field formed in a direction orthogonal to one direction of the flow channel 110. The particles having positive electric charge are moved by electrophoresis, and electrophoretic fluidity The distance traveled depends on the distance. The electrophoretic fluidity may vary depending on the particle size, amount of charge, buffer composition, and the like.

Flows through the plurality of discharge units 150 connected to the other end of the flow channel 110 so that the biological sample particles separated according to the electrophoretic fluidity passing through the electric field forming unit 140 are isolated from each other, . 2 and 3, one end of each of the discharge units 150 may have a different height from the flow channel 110, that is, That is, the direction orthogonal to the longitudinal direction of the flow channel 110, as shown in FIG.

Referring to FIG. 3, the electric field forming unit 140 may gradient the pH of the biological sample flowing along the flow channel 110 or form an electric field inside the flow channel 110, as described above. The electric field forming unit 140 includes a positive ion supplying unit 146, an anion supplying unit 143, and a power supplying unit 149.

The cation supply unit 146 is connected to one side of the flow channel 110 and supplies positive ions to the flow channel 110. The first and second storage units 144 and 144, And a negative charge polymer 145 that selectively transmits only positive ions among the ions contained in the first electrolyte. At this time, the first electrolyte preferably contains hydrogen ions (H + ), and more preferably HCl.

The negative charge polymer 145 selectively permeates only cations among the ions (that is, cations and anions) contained in the electrolyte, for example, poly-AMPS (2-acrylamido-2-methyl-1-propanesulfonic acid) poly-SS (styrene sulfonate) is preferably used. The negative charge polymer 145 is disposed between the first reservoir 144 and the flow channel 110 and transmits only the positive ions included in the first electrolyte, for example, hydrogen ions H + , to the flow channel 110 .

The anion supply unit 143 is connected to the flow channel 110 so as to be disposed on the opposite side of the cation supply unit 146 with the flow channel 110 interposed therebetween to supply negative ions to the flow channel 110 And may include a second storage unit 141 storing a second electrolyte, and a positive charge polymer 142 selectively transmitting only anions among the ions contained in the second electrolyte. At this time, the second electrolyte preferably includes a hydroxide ion (OH < - & gt ; ). More preferably, the second electrolyte is KOH.

The positively charged polymer 141 selectively permeates only anions among ions contained in the electrolyte (that is, cations and anions). For example, poly-DADMAC (diallyldimethylammonium chloride) may be used. The positively charged polymer 142 is disposed between the second reservoir 141 and the flow channel 110 and transmits only anions included in the second electrolyte, for example, hydroxide ions OH - to the flow channel 110 .

The power supply 149 supplies the ions from the cation supply part 146 and the anion supply part 143 to the flow channel 110 and applies power to form an electric field. The power supply unit 149 includes a pair of electrodes 147 and 148 and the pair of electrodes 147 and 148 are connected to the first storage unit 144 and the second storage unit 141, .

Hereinafter, the process of separating the biological sample using the microfibrillator separation apparatus 100 configured as described above will be described below.

First, when the buffer and the biological sample are injected through the first inlet and the second inlet connected to one end of the flow channel 110, the flow rate of the buffer is adjusted so that the biological sample is separated from the buffer along one side wall of the flow channel And flows in parallel.

When power is supplied through the power supply unit, positive ions are supplied to the flow channel from the positive ion supply unit of the electric field forming unit 140, and negative ions are supplied to the flow channel from the negative ion supply unit. As a result, An electric field is formed.

In this state, in the process of passing the biological sample flowing along one side wall of the flow channel through the electric field forming part 140, the particles (proteins or microvesicles contained in the biological sample) Becomes cations and flows in the direction of the formed electric field. At this time, the moving distance varies depending on the electrophoretic fluidity which varies depending on factors such as the particle size, charge amount, buffer composition, and the like. Accordingly, the particles are separated from each other, and the separated particles passing through the electric field forming unit 140 are isolated while flowing through a plurality of discharge units 150 connected to the other end of the flow channel, finally separated.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, It is evident that it is possible to modify or modify it by the owner.

100: microfibrillar separator 110: flow channel
120: first inlet 130: second inlet
140: electric field forming part 141: negative ion supplying part
142: positive charge polymer 143: negative ion supply part
144: first storage unit 145: negative charge polymer
146: Cation supply part 147: Electrode
148: Electrode 149: Power supply
150:

Claims (10)

A flow channel in which the biological sample and the buffer are separated and flow side by side along one direction;
A first injection port connected to a first path at one end of the flow channel to inject the buffer;
A second injection port connected to a second path at one end of the flow channel to inject the biological sample;
An electric field forming unit for forming an electric field in the flow channel in a direction orthogonal to the one direction so that the biological sample flowing along the flow channel is separated according to electrophoretic fluidity; And
And a plurality of discharging portions connected to the other end of the flow channel so that the separated samples are separated and flowed according to the electrophoretic fluidity.
The method according to claim 1,
The biological sample
Protein, micro-endoplasmic reticulum, or a mixture thereof.
The method according to claim 1,
The flow channel
Wherein the flow rate of the buffer to be injected is adjusted so that the biological sample flows along one side wall of the flow channel separately from the buffer and flows side by side.
The method according to claim 1,
The flow rate of the injected buffer
Wherein the flow rate of the biological sample to be injected is 1 to 20 times the flow rate of the biological sample to be injected.
The method according to claim 1,
The electric field forming unit includes:
A cation supply part connected to one side of the flow channel and supplying cations to the flow channel;
An anion supply unit connected to the flow channel so as to be disposed on the opposite side of the cation supply unit with the flow channel interposed therebetween, the anion supply unit supplying an anion to the flow channel; And
A power supply unit having a pair of electrodes, each of which is provided to the positive ion supply unit and the negative ion supply unit, respectively; Wherein the microfibrillar separator is a microfibrillar separator using electrophoresis.

The method of claim 5,
The electric field forming unit
The particles included in the passing biological sample are positively charged by positive ions supplied from the cation supply unit,
Wherein the microembossed particles having a positive charge are separated by an electrophoretic fluidity while flowing through an electric field formed in a direction orthogonal to the one direction.
The method of claim 5,
The cation supply part
A first storage unit storing a first electrolyte; And
A negative charge polymer that selectively transmits only positive ions among the ions included in the first electrolyte; Wherein the microfibrillar separator is a microfibrillar separator using electrophoresis.
Claim 5
The anion supply unit
A second reservoir storing a second electrolyte;
A positive charge polymer that selectively transmits only anions among the ions contained in the second electrolyte; Wherein the microfibrillar separator is a microfibrillar separator using electrophoresis.
The method of claim 8,
The first electrolyte
Hydrogen ions (H & lt ; + & gt ; ),
The second electrolyte
And a hydroxide ion (OH < - & gt ; ).
The method of claim 9,
The first electrolyte
HCl,
The second electrolyte
KOH. ≪ / RTI >
KR1020150066864A 2015-05-13 2015-05-13 Apparatus for separating fine endoplasmic reticulum using electrophoresis KR20160133826A (en)

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Cited By (1)

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CN110479393A (en) * 2019-08-24 2019-11-22 德运康明(厦门)生物科技有限公司 A kind of programmable micro-fluidic chip surface modification of and coloration instrument

Citations (1)

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Publication number Priority date Publication date Assignee Title
KR20050119128A (en) 2003-03-19 2005-12-20 알파 와슬만, 인크. Separation and accumulation of subcellular components, and proteins derived therefrom

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050119128A (en) 2003-03-19 2005-12-20 알파 와슬만, 인크. Separation and accumulation of subcellular components, and proteins derived therefrom

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110479393A (en) * 2019-08-24 2019-11-22 德运康明(厦门)生物科技有限公司 A kind of programmable micro-fluidic chip surface modification of and coloration instrument
CN110479393B (en) * 2019-08-24 2021-11-12 苏州德运康瑞生物科技有限公司 Programmable micro-fluidic chip surface modification dyeing instrument

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