CN1477391A - Method for sedparating bialogical macromolecule by using two-dimensinal or multi-dimensinal capillary electrophoresis and its used interface - Google Patents
Method for sedparating bialogical macromolecule by using two-dimensinal or multi-dimensinal capillary electrophoresis and its used interface Download PDFInfo
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Abstract
The present invention relates to a method for separating biological macromolecular component by using two-dimensional or multidimensional capillary electrophoresis and its interface. The biological macromolecular component undergone the process of one-dimensional capillary electrophoresis separation can be passed through the interface and fed into two-dimensional capillary electrophoretic separation operation to implement two-dimensional capillary electrophoresis, the small molecular substance can be diffused and freely permeated through the hollow fibre in the interface and can obtain balance in it, and when the described biological macromolecular component is flowed through the interface, the buffer solution containing low molecular substance can be on-line added/removed into the hollow fibre in the interface.
Description
Technical field
The present invention relates to capillary electrophoresis technique, method and the used interface of specifically a kind of two dimension or multidimensional capillary electrophoresis separation biomacromolecule.
Background technology
Capillary Electrophoresis (CE) be a class easy, separate efficiently, analytical approach.Compare with the disk electrophoresis method, have fast, amount of samples is few, be easy to plurality of advantages such as robotization, in the application that has obtained aspect the compartment analysis of life active compounds such as protein, antibody, clinical sample more and more widely.
Imitating although capillary electrophoresis method has higher post, generally is a dimensional pattern, because the restriction of applied voltage has restricted the separating column length that is adopted, thereby has also limited the peak capacity that actual post is imitated and may be reached.In addition, the meticulous internal diameter of capillary column makes that its spectral detection light path is too small, and detection sensitivity is produced a very large impact.These restrictions and influence are particularly outstanding in complex biochemical sample, Analysis of environmental samples.Therefore development has than the Capillary Electrophoresis instrument and the method for peak capacity and higher detection sensitivity are of great significance for further widening its application and research field greatly.
(document 1:Gidings L C, Unified Seperation Sciecc, John Wilev﹠amp such as Gidding; Sons, New York) at first propose two kinds of clastotypes in conjunction with constituting the thought that the multidimensional system is used for the separation of complex sample.Adopt the multidimensional isolation technics can greatly improve the peak capacity of system, also can be more easily and highly sensitive detecting device coupling, finish the compartment analysis task of complex sample better.At present, two-dimensional gas chromatography is commercialization, and successfully is used for the separation of refinery gas etc., has reached extraordinary effect.Two-dimensional gel electrophoresis is the important tool of present protein groups research, and the work of trial property has been carried out in the research of two-dimensional liquid chromatography also someone.The feature that transports according to fluid in the kapillary, the method that adopts TURP to change is carried out technology existing application in capillary chip separates that multidimensional is separated, especially in human genome research, obtaining development preferably, (document 2: Sun Yue such as Guan Yafeng, Guan Yafeng, analytical chemistry, 25 (7), 745-749,1997; Document 3: Sun Yue, Guan Yafeng, chromatogram, 15 (2), 106-109,1997) once attempted on chip, realizing the combination of different mode CE.But because the separation column length of chip is too short, can not satisfy the requirement for the complex sample peak capacity, this method also is unsuitable for samples such as big molecular activity albumen simultaneously, low its compartment analysis to trace components that also limited of detection sensitivity.
Realized in recent years that CE and other separate modes comprised coupling (document 4:Hooker, the Thomas F. of liquid chromatography (LC); Jeffery, Dorothea J.; Jorgenson, James W, Cherm.Anal., 146,581-612 1998; Document 5:Hooker, T.F., Jorgenson, J.W., Anal.Chem.1997,69,4134-4142; Document 6:Moore, A.W.Jr., Jorgenson, J.W., Anal.Chem.1995,67,3456-3463; Document 7:Liu, Zaiyou; Lee, Milton L.J.Microcolumn Sep., 12 (4), 241-254 2000; Document 8:Manabe, Takashi, Electrophoresis, 21 (6), 1116-1122,2000; Document 9:Neuhoff, Volker, Electrophoresis, 21 (1), 3-11,2000; Document 10:Bushey, Michelle M.; Jorgenson, James W, Anal.Chem., 62 (10), 978-84,1990; Document 11:Tang, Qing; Harrata, A.Kamel; Lee, Cheng S., Anal.Chem., 69 (16), 3177-3182,1997), obtained and the similar result of conventional two-dimensional electrophoresis spectrogram, done preliminary trial for the two-dimentional coupling of Capillary Electrophoresis-Capillary Electrophoresis also someone, electronic or mechanical switching realizes that sample enters another capillary post by a capillary post to these methods based on intercolumniation, and peak capacity is greatly improved.Two dimensional separation is used for the complex biological sample separation at Capillary Electrophoresis, analysis has very important significance, and people notice the advantage that multidimensional is separated gradually recently, and expection will become research focus new in the separation science.But so far, the coupling between the two dimension of high post number or multidimensional Capillary Electrophoresis and different mode yet there are no report.
Summary of the invention
In order to overcome above-mentioned deficiency, the purpose of this invention is to provide a kind of post is imitated, peak capacity is higher, be suitable for the complex sample requirement two dimension or method and the used interface of multidimensional capillary electrophoresis separation biomacromolecule, it has the advantages that high post number, sample introduction sample reach coupling between different capillary electrophoretics continuously.
To achieve these goals, technical scheme of the present invention is: finish the artifact macromolecular components interface of flowing through in the first dimension capillary electrophoresis separation and enter the second dimension capillary electrophoresis separation operation, realize two-dimensional capillary electrophoresis, and small-molecule substance can pass through to spread the hollow fiber that freely sees through in the interface, and reaches balance at it;
The buffer solution that contains low molecular weight substance is added/removed to described biomacromolecule component stream in the hollow fiber in the alignment interface when interface, realize dynamic flushing simultaneously at the buffer solution interface of flowing through continuously, give on the capillary column that separating or that separated simultaneously and apply voltage simultaneously, to adjust the chargeding performance of the biomacromolecule that from hollow fiber, passes through;
To enter down the one dimension capillary column by next interface by the biomacromolecule component of above-mentioned processing and further separate, thereby realize the multidimensional Capillary Electrophoresis, same small-molecule substance can freely see through hollow fiber by diffusion, and the outer within it balance that reaches; The buffer solution that contains low molecular weight substance is added/removed to described biomacromolecule component stream in the hollow fiber in the alignment interface when interface, realize dynamic flushing simultaneously at the buffer solution interface of flowing through continuously, give on the capillary column that separating or that separated simultaneously and apply voltage simultaneously, to adjust the chargeding performance of the biomacromolecule that from hollow fiber, passes through; Described pressuring method is: only apply voltage on the first dimension capillary column when the first dimension capillary electrophoresis separation, when first and second dimension capillary electrophoresis separation, on first and second dimension capillary column, apply voltage simultaneously, by that analogy, promptly give on the capillary column that separates or separated, to apply voltage simultaneously; The described Control of Voltage that applies is in 5000～30kV scope; The described interface that adds/remove the buffer solution that contains low molecular weight substance in the hollow fiber of alignment biomacromolecule component stream warp does not comprise the first dimension capillary sample inlet end interface; Described low molecular weight substance is surfactant, salt; Described capillary electrophoresis separation pattern can be respectively capillary isoelectric focusing electrophoresis (CIEF), capillary zone electrophoresis (CZE), kapillary electrodynamics chromatogram (MEKC) or capillary electric chromatogram (CEC);
Adopt electroosmotic flow to drive the two dimensional separation system, the electrode of tieing up the endpiece of capillary column by the first dimension capillary column sample introduction termination electrode, interface electrode and second constitutes three-electrode system, keeps the potential gradient in the bidimensional capillary post, reaches the separating effect of optimization; Adopt pressure-actuated detachment process, by the interface between the sealed hair capillary column, keep the hollow fiber external and internal pressure identical, apply 0.001 to 10MPa pressure at the first dimension capillary column entrance point then, the biomacromolecule component that first dimension is separated enters the second dimension capillary column; Adopt electroosmotic flow to drive the multidimensional piece-rate system, the electrode formation multi-electrode system of the endpiece by preceding one dimension capillary column sample introduction termination electrode, interface electrode and following one dimension capillary column is kept the potential gradient in the multidimensional capillary column, reaches the separating effect of optimization; Adopt pressure-actuated detachment process, by the interface between the sealed hair capillary column, keep the hollow fiber external and internal pressure identical, apply 0.001 to 10MPa pressure then at preceding one dimension capillary column entrance point, the biomacromolecule component that one dimension separates before making enters down the one dimension capillary column;
Its used interface has the interface repository body, on the interface repository body, be provided with electrode socket, inlet, outlet, hollow fiber is installed between the connector at interface repository body two ends, capillary column is complementary to seal by connector and the inside and outside footpath of the port at hollow fiber two ends and docks, be inserted with electrode on the electrode socket and stretch into the interface repository body, and leave distance between the hollow fiber.
The present invention has following advantage:
1. the characteristics that have coupling between different capillary electrophoretics.The present invention has semipermeable character according to the hollow fiber interface, when holding back biomacromolecule, allow micromolecule solute, solvent freely to pass through, can in piece-rate system, add for example yin, yang ionic surface active agent of micromolecule composition by it, also can from system, slough unnecessary surfactant or salinity by the way of continuous sample introduction and flushing, thereby the independent assortment of convenient various CE patterns, promptly the facility of multiple clastotype connects.
2. post is imitated high.The hollow fiber that the present invention adopts and the internal diameter of capillary column mate mutually, realize that no dead volume connects, and biomacromolecule is moved continuously along split tunnel, and not to the sample cell diffusion, no component are lost diluted sample and heap problem in phenomenon, the no sample introduction process; Adopt the present invention to carry out Capillary Electrophoresis and can realize that two-dimensional system moves respectively under 0.001 to 10MPa pressure easily by keeping the pressure of the first dimension capillary column sample introduction end, the second dimension capillary column endpiece and interface; Can realize respectively in the multidimensional system that also pressure-driven, electroosmotic flow driving or pressure, electroosmotic flow drive simultaneously.In addition, because interface can seal and bearing certain pressure, can suppress to produce in the separating column bubble.
3. can satisfy detection requirements such as mass spectrum.Interface of the present invention is provided with the function of solvent exchange, can realize the online displacement of buffer solution, can conveniently realize operations such as protein denaturation, renaturation, enzymolysis in interface arrangement, so the present invention can be used for satisfying the needs of detection system to buffer solution.
4. easy to operate.The present invention can adopt single electrophoretic, also can coupling between different electrophoretics, also can adopt electrode common technology and electroosmotic flow adjustment technology, and only use a high-voltage power supply can realize two dimension, multidimensional capillary electrophoresis separation.
5. applied range.Interface arrangement of the present invention selects molecular weight to hold back different hollow fibers, can handle the biomacromolecule sample of different molecular weight, as oligonucleotide, polypeptide, protein and DNA etc.; Also can be used for the reactions such as post column derivatization after first dimension is separated, interface arrangement of the present invention uses so also can be used as reactor owing to play solvent exchange.
6. peak capacity is higher.Interface of the present invention can be realized the independent assortment of different capillary electrophoretics, does not therefore restrict the separating column length that is adopted; Again, the realization of two dimension or multidimensional capillary electrophoresis technique, the complex component that capillary electrophoretic can not be handled fully can reach thorough separation and sign by the coupling of multidimensional capillary electrophoresis technique, shown in Fig. 3,4, the sample introduction that comprises the multidimensional capillary column that the bidimensional capillary post is later, be each spectrum peak of separating based on the capillary column of its front, so post is imitated and will be greatly improved (above 10
6The number of plates/m), peak capacity also can further improve simultaneously.
Description of drawings
Fig. 1 is an interface arrangement structural representation of the present invention.
Fig. 2 is capillary isoelectric focusing (CIEF) spectrogram of haemoglobin in the one embodiment of the invention.
Fig. 3 is the capillary isoelectric focusing-capillary gel electrophoresis (CIEF-CGE) of haemoglobin in the one embodiment of the invention.
Fig. 4 is the capillary isoelectric focusing-capillary gel electrophoresis-capillary zone electrophoresis (CIEF-CGE-CZE) of haemoglobin in the one embodiment of the invention.
Embodiment
Below in conjunction with embodiment and accompanying drawing the present invention is described in further detail.
Embodiment 1
Experiment condition:
Capillary column (Hebei Yongnian sharp foreign chromatogram device company limited, 50 μ m I.D., 375 μ m O.D.), carrier ampholyte Pharmalyte (pH3.0-10.0, BioChemika, Switzerland), the biomacromolecule component is a haemoglobin, be the biochemical institute in Shanghai product, other (its reagent name of the present invention) reagent are pure for analyzing.
Capillary electrophoresis apparatus is the TriSep of Unimicro Technology
TM-2000GV type comprises a Data Module wavelengthtunable ultraviolet-visible detecting device, the continuous adjustable high-voltage DC power supply of CEC control module; Data aggregation and spectrogram are handled and are adopted according to the Echrom98 of Lyntech Corporation (US) 10177 South 77th East Avenue Tulsa, Oklahoma 74133 U.S. workstation and Echrom98 V2.0 software.
The present invention realizes that the special purpose interface of the method for two dimension or multidimensional capillary electrophoresis separation biomacromolecule has interface repository body 2, on interface repository body 2, be provided with electrode socket 4, inlet 5, outlet 6, hollow fiber 3 is installed between the connector 7 at interface repository body 2 two ends, capillary column l is complementary to seal by the connector 7 and the inside and outside footpath of port at hollow fiber 3 two ends and docks, be inserted with electrode on the electrode socket 4 and stretch into interface repository body 2, and leave distance between the hollow fiber 3; Described interface repository body 2 is quartz glass, pottery or polymkeric substance, and present embodiment is a quartz glass.
The present invention realizes the method for two dimension or multidimensional capillary electrophoresis separation biomacromolecule, and its concrete operations step is as follows:
First dimension adopts capillary isoelectric focusing (CIEF) method, and buffer solution is 30mmol/Ltris-HCl (containing 2% (v/v) Pharmalyte pH3-10), and anodal electrolytic solution is 30mmol/LH
3PO
4, negative pole electrolytic solution is 30mmol/L NaOH, whole post sample introduction, second dimension adopts capillary gel electrophoresis (CGE) method, the third dimension adopts capillary zone electrophoresis (CZE) method, used kapillary column length is 25cm, effective length 15cm.The experiment applied voltage is 8kV.
Give during capillary electrophoresis separation first to tie up on the capillary column and apply 8kV voltage in first dimension, after separation is finished, with inlet 5, outlet 6 continuous (flow velocity 2mL/mins) the flushing interface of 30mmol/L tris-HCl buffer solution (containing the 0.2%w/v lauryl sodium sulfate) by interface arrangement, be implemented in to add in the hollow fiber 3 in the alignment interface and contain low molecular weight substance (0.2%w/v lauryl sodium sulfate), (part) removes low molecular weight substance (carrier ampholyte Pharmalyte) simultaneously.Destroyed through the pH gradient that isoelectric focusing (IEF) process forms, the migration of enforcement negative pole, the biomacromolecule component enters the second dimension capillary column (wherein be full of in advance the described 0.2%w/v of containing lauryl sodium sulfate contain low molecular weight substance buffer solution) through interface, all apply 8kV voltage on first and second dimension capillary column this moment, when the biomacromolecule component enters the second dimension capillary column, all saturated and be with negative charge by lauryl sodium sulfate institute, under the driving of electric field by negative pole to anodal swimming, realize the CIEF-CGE two-dimensional capillary electrophoresis;
After the second dimension capillary electrophoresis separation is finished, interface with 30mmol/L tris-HCl buffer solution continuous (flow velocity 2mL/min) the flushing second dimension kapillary two ends, realize that online (part) removes low molecular weight substance (lauryl sodium sulfate) and apply 8kV voltage simultaneously simultaneously on first, second and third dimension kapillary, the biomacromolecule component of coming out from the second dimension capillary column enters third dimension capillary column through interface, under the driving of normal electrical seepage flow, protein component to the negative pole swimming, is realized the three-dimensional Capillary Electrophoresis of CIEF-CGE-CZE by positive pole smoothly.
Its experimental result:
As shown in Figure 2, the biomacromolecule component is a haemoglobin process capillary isoelectric focusing electrophoresis, is separated into 5 peaks by the isoelectric point difference.
As shown in Figure 3, after process capillary isoelectric focusing electrophoretic separation is 5 peaks, drive through chemistry, each peak once enters the second dimension kapillary by interface, successively further separate according to the capillary gel electrophoresis pattern, each peak all further is separated into different peaks.Realized the coupling of CIEF-CGE different mode smoothly.Because each spectrum peak that CGE separates with CIEF is basic sample introduction, so the post effect will be greatly improved (above 10
5The number of plates/m).
As shown in Figure 4, the component of similarly separating through two-dimentional CIEF-CGE further is separated into more peak by capillary zone electrophoresis.Realized the coupling of CIEF-CGE-CZE different mode smoothly.Because each spectrum peak that CZE separates with CGE is basic sample introduction, so the post effect will be further enhanced (above 10
6The number of plates/m), peak capacity is further enhanced simultaneously.
Embodiment 2
Difference from Example 1 is: the online buffer solution that contains low molecular weight substance that removes is that salt is identical with embodiment 1, when implementing the second dimension capillary gel electrophoresis separation (voltage 8kV), making alive not on the first dimension capillary column replaces the pressure of 0.1Mpa, is used for driving the spectrum peak; Same when implementing the gel electrophoresis of third dimension capillary column and separate (voltage 8kV), making alive not on first and second dimension capillary column replaces the pressure of 0.1Mpa, is used for driving the spectrum peak, can obtain similar result.
Relevant comparative example
The multi-mode multidimensional analysis schematic diagram that the gel chromatography/liquid chromatography of people such as Jorgenson design/Capillary Electrophoresis is formed adopts the T-valve interface, can not continued operation, in addition to high performance liquid chromatogram and fast the Capillary Electrophoresis interconnection technique have relatively high expectations.The sample size of Capillary Electrophoresis very little (receive upgrading nL) on the one hand, in order to reach purpose rapidly and efficiently, to efficiently the repeating to introduce and require very highly of liquid chromatography effluent, so they adopt laser beam control technology sample introduction.To feed electrophoresis continuously rough for sample in the analytic process, and in the porch sample that flows into is taken off with the laser beam branch.This interfacing is unsuitable for the coupling of different capillary electrophoretics, and the buffer solution of electrophoresis also is subjected to the influence of liquid phase chromatogram condition simultaneously.
(document 12: Zhang Xiangmin etc. such as Zhang Xiangmin, significant problem in the traditional Chinese medicine complex system is inquired into, chief editor Wang Xiaoru, publishing house of Xiamen University, 1998) designed a kind of interface, be used for the connection of liquid chromatography/Capillary Electrophoresis two-dimentional system, liquid phase capillary column and electrophoretic column are fixed on the coaxial cable, the two ends distance is about 150pm, and add a sleeve pipe movably at the liquid-phase chromatographic column end, pass through electrophoretic buffer two pillar tie point vertical direction, and maintain certain quantity solution by the slit between two pillars.When sleeve pipe moves to slit, the flowing of shield solution, liquid chromatography flows out component and begins to pile up at electrophoresis kapillary column cap in the slit, adds that voltage just can carry out the electrophoresis sample introduction; Sample introduction finishes, and mobile sleeve pipe is to original position, and solution is taken away the liquid phase effluent by slit again.Therefore, just can be interrupted the operation of electrophoresis sample introduction by mobile sleeve pipe.This sampling technique has two aspect advantages: reduce the dilution of sample effect and improve sensitivity and repeatability: improve sample introduction speed and be easy to automatic control.These advantages are quite important to the performance that improves the bidimensional separation interface.The coupling of different capillary electrophoretics but this interfacing still is unrealized.
Hooker etc. have developed new interfacing method, adopt transparent material and valve handoff technique control sample introduction process, this method has universality, but the use of valve can cause sample introduction discontinuous, there are diluted sample and heap in the sample introduction process simultaneously, need problems such as effluent is more.
Chinese patent application (application number: 01127918.4) disclose a kind of stereo multi-dimensinal multi-mode capillary electrophoresis method and isolated plant, wherein interface is a kind of cross type structure of little dead volume, be used for the switching of CE/CE two dimensional separation system, its method is to switch by voltage or additive method moving phase is switched between different streams, realize CE/CE two dimensional separation system, superpotential or additive method changing method all do not cause component to be lost and the uncontinuity of sample introduction easily.
Above-mentioned interface does not all have the function of solvent exchange, can not adjust buffer solution according to actual conditions when separating, and the step of going forward side by side joint is by pH value, ionic strength, the polarity of the separated component of interface.
Claims (10)
1. the method for two dimension or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: finish the artifact macromolecular components interface of flowing through in the first dimension capillary electrophoresis separation and enter the second dimension capillary electrophoresis separation operation, realize two-dimensional capillary electrophoresis, and small-molecule substance can pass through to spread the hollow fiber that freely sees through in the interface, and reaches balance at it;
The buffer solution that contains low molecular weight substance is added/removed to described biomacromolecule component stream in the hollow fiber in the alignment interface when interface, realize dynamic flushing simultaneously at the buffer solution interface of flowing through continuously, give on the capillary column that separating or that separated simultaneously and apply voltage simultaneously, to adjust the chargeding performance of the biomacromolecule that from hollow fiber, passes through.
2. press the method for described two dimension of claim 1 or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: will enter down the one dimension capillary column by next interface by the biomacromolecule component of above-mentioned processing and further separate, thereby realize the multidimensional Capillary Electrophoresis, same small-molecule substance can freely see through hollow fiber by diffusion, and the outer within it balance that reaches; The buffer solution that contains low molecular weight substance is added/removed to described biomacromolecule component stream in the hollow fiber in the alignment interface when interface, realize dynamic flushing simultaneously at the buffer solution interface of flowing through continuously, give on the capillary column that separating or that separated simultaneously and apply voltage simultaneously, to adjust the chargeding performance of the biomacromolecule that from hollow fiber, passes through.
3. press the method for claim 1 or 2 described two dimensions or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: described pressuring method is: only apply voltage on the first dimension capillary column when the first dimension capillary electrophoresis separation, when first and second dimension capillary electrophoresis separation, on first and second dimension capillary column, apply voltage simultaneously, by that analogy, promptly give on the capillary column that separates or separated, to apply voltage simultaneously; The described Control of Voltage that applies is in 5000～30kV scope.
4. by the method for claim 1 or 2 described two dimensions or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: the described interface that adds/remove the buffer solution that contains low molecular weight substance in the hollow fiber of alignment biomacromolecule component stream warp does not comprise the first dimension capillary sample inlet end interface; Described low molecular weight substance is surfactant, salt.
5. by the method for claim 1 or 2 described two dimensions or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: described capillary electrophoresis separation pattern can be respectively capillary isoelectric focusing electrophoresis, capillary zone electrophoresis, kapillary electrodynamics chromatogram or capillary electric chromatogram.
6. press the method for described two dimension of claim 1 or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: adopt electroosmotic flow to drive the two dimensional separation process, the electrode of tieing up the endpiece of capillary column by the first dimension capillary column sample introduction termination electrode, interface electrode and second constitutes three-electrode system, keep the potential gradient in the bidimensional capillary post, reach the separating effect of optimization.
7. press the method for described two dimension of claim 1 or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: adopt pressure-driven two dimensional separation process, by the interface between the sealed hair capillary column, keep the hollow fiber external and internal pressure identical, apply 0.001 to 10MPa pressure at the first dimension capillary column entrance point then, the biomacromolecule component that first dimension is separated enters the second dimension capillary column.
8. press the method for described two dimension of claim 2 or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: adopt electroosmotic flow to drive the multidimensional detachment process, the electrode of the endpiece by preceding one dimension capillary column sample introduction termination electrode, interface electrode and following one dimension capillary column constitutes multi-electrode system, keep the potential gradient in the multidimensional capillary column, reach the separating effect of optimization.
9. press the method for described realization two dimension of claim 2 or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: adopt the detachment process of pressure-driven multidimensional, by the interface between the sealed hair capillary column, keep the hollow fiber external and internal pressure identical, apply 0.001 to 10MPa pressure then at preceding one dimension capillary column entrance point, the biomacromolecule component that one dimension separates before making enters down the one dimension capillary column.
10. method used interface by claim 1 or 2 described two dimensions or multidimensional capillary electrophoresis separation biomacromolecule, it is characterized in that: have interface repository body (2), on interface repository body (2), be provided with electrode socket (4), inlet (5), outlet (6), hollow fiber (3) is installed between the connector (7) at interface repository body (2) two ends, in the port of capillary column (1) by connector (7) and hollow fiber (3) two ends, external diameter is complementary and seals butt joint, be inserted with electrode on the electrode socket (4) and stretch into interface repository body (2), and leave distance between the hollow fiber (3).
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