CN104297491B - Protein chromatographic electrophoresis and in-situ chemical imprinting and immune imaging device and method - Google Patents

Abstract

The invention discloses protein chromatographic electrophoresis and an in-situ chemical imprinting and immune imaging device and method. The device comprises chromatographic electrophoresis modules, first solution tanks, a chromatographic electrophoresis separation plate with multiple electrophoresis channels, active chromatographic media and an LED fluorescent light source, wherein two ends of the chromatographic electrophoresis separation plate are respectively connected with the first solution tank and the chromatographic electrophoresis module; the active chromatographic media are contained in each electrophoresis channel and comprise microparticles which are composed of surface-activated ion exchange resin, affinity chromatography fillers and reverse phase chromatography fillers or are non-porous silica microparticles or porous glass microparticles or microparticles made from high molecular materials; and diazirine coupling molecules are dielectrically bonded on the surfaces of the microparticles. A whole process of various chromatographic electrophoresis separation and in-situ chemical imprinting and immune imaging of protein can be finished within two hours. The device and the method disclosed by the invention are high in sensitivity, distinguishability and repeatability while the characteristics of traditional protein chromatography and electrophoresis are maintained.

Description

Protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging device and method

Technical field

The present invention relates to medical science, more particularly, to a kind of protein chromatographic electrophoresis and its in-situ chemical trace and exempt from Epidemic disease imaging device and method.

Background technology

Traditional immune protein imaging, or claim Western, the polyacrylamide gel electrophoresis including albumen separates, coagulates In glue, the transfer trace of albumen, the immune detection relying on antigen antibody interaction and protein immunization imaging analysis four are important Technology forms, and is the most frequently used protein analysis method in modern life science and clinic study.Traditional Western analysis Need successively to use multiple instrument and equipments, complex operation step, technical requirements are high, it usually needs two day time just can complete, no Method realizes automation, and the result obtaining usually changes because of the difference of instrument, reagent, method and operator.Due to tradition Western analysis is made up of four complicated technology, it usually needs four kinds of instrument and equipments are completing：Protein electrophoresis instrument, albumen turn Film instrument, protein immunoblot instrument and protein immunization imager.Wherein albumen polyacrylamide gel electrophoresis complex operation it is impossible to To be imaged on automatic mode first-class compatible with protein delivery trace, protein immunization.So, going back neither one instrument and equipment can To complete four complex technology operations of traditional Western analysis.Each individual event instrument and equipment feelings of composition traditional Western analysis Condition is as follows：1) traditional protein electrophoresis instrument：I.e. albumen by size, the property such as electric charge separately.Traditional protein electrophoresis has SDS-PAGE Vertical electrophoresis apparatus and isoelectric focusing electrophoresis instrument.Wherein SDS-PAGE vertical electrophoresis apparatus are very universal, and isoelectric focusing electrophoresis instrument with Solid phase isoelectric focusing adhesive tape (IPG) electrophoresis system of U.S. Bio-rad Bole and U.S.'s Thermo thermoelectricity, and Agilent Based on the offgel Liquid isoelectric focusing electrophoresis apparatus of Agilent.The critical defect of both electrophoresis is that all of SDS-PAGE hangs down Straight electrophoresis apparatus, due to using polyacrylamide gel medium, all cannot accomplish full-automation；All of isoelectric focusing electrophoresis, by In using IPG adhesive tape, real time imagery all cannot be realized.2) albumen transferring film instrument presses its point the albumen in polyacrylamide gel Cloth is transferred to exactly on nitrocellulose filter or pvdf membrane, forms Western blot.Although a lot of professional persons still continue to use The transfer liquid of traditional-handwork inhales profit method, and electrophoretic transfer has been widely used.Electrophoretic transfer must be printed using protein delivery Mark instrument.The market of the instrument of protein electrophoresis transfer at present is more chaotic, does not have quality standard.3) protein immunoblot instrument：I.e. in nitric acid Antigen antibody interaction is completed on cellulose membrane or pvdf membrane.Due to two kinds of antibody will be used, and multiple film to be completed Protection and washing, automation process is comparatively laborious.But exactly process is loaded down with trivial details, people are very high to the demand of automation.Over the years, It is proposed full-automatic protein immunoblot instrument respectively both at home and abroad.The full-automatic Diagnosis of Sghistosomiasis of B20 type of such as Britain Bee Robotics The XD236 automatic westem blot instrument that mark instrument, ProfiBlot48, AutoBlot36 of U.S. TECAN and domestic upper Hisoon reach.But Due to expensive, using not being very universal.4) immune imager：I.e. on nitrocellulose filter or pvdf membrane with antibody phase The albumen colour developing of interaction in situ imaging.According to the difference of antibody labeling method and colour developing principle, immune imager can be divided into Chemiluminescence imaging instrument and phosphorimager.Wherein, with the Odyssey near-infrared laser imager of LICOR, THERMO The LAS-4000 fluorescence of typhoon Multifunction fluorescent scanner and FUJI and chemiluminescence imaging instrument are representative.At present, main Will be based on import equipment.So, there are a lot of problems first in traditional Western analysis, traditional Western analysis needs successively Using multiple instrument and equipments, complex operation step, technical requirements are high, it usually needs two day time just can complete all to analyze, and catches up with Do not go up the paces of Development of Modern Science；And, the result of traditional Western analysis is usually because of instrument, reagent, method and operator Difference and change it is impossible to meet the requirement to protein quantification in biomedicine；Four technology compositions of traditional Western analysis Part cannot be compatible on automatic mode, there is presently no an instrument and equipment and can complete traditional Western analysis All processes；Finally, Western analysis method clinically has extensive demand, but present operation is all open, The contact that operating personnel are with detection sample cannot be avoided, clinical position may be threatened by infectiousness sample.

With respect in traditional Western analysis, with polyacrylamide gel as electrophoretic medium, Capillary Electrophoresis is with capillary Manage as split tunnel, the trace of albumin separation method with free solution as medium.Generally, Capillary Electrophoresis uses internal diameter is 25- 100 μm of elasticity (polyimides) coating vitreous silica capillary, it is not necessary to solid dielectric in addition to electrophoretic buffer.So, Volume is little, rapid heat dissipation, can bear high electric field (100-1000V/cm), easily realizes automation.Recently, with Capillary Electrophoresis as base The full-automatic protein immunization analytical instrument of plinth has been come out, and can complete trace, the albumen Western of even tens cells divides Analysis.The advantage of Capillary Electrophoresis protein immunization analysis：1) efficient, plate number is between 105-107 piece/m；2) quick, typically exist Complete in more than ten minutes to separate；3) micro, the sample volume needed for sample introduction is nanoliter level；4) automate, be to automate journey at present Spend higher separation method.The shortcoming of Capillary Electrophoresis protein immunization analysis is 1) because sample size is few, thus preparative capacibility is poor, In particular for the amount required for during further mass spectral analysis, being often unable to reach；2) because capillary diameter is little, make light path too Short, detect or image sensitivity relatively low (as ultraviolet absorption spectroscopy)；3) because electroosmosis is different because sample forms, albumen divides From poor reproducibility；4) instrument and equipment is expensive, and special capillary consumables cost is too high, limits its popularization and application.Contrast it Under, lacking in the advantage exactly Capillary Electrophoresis protein immunization analysis system of traditional Western analysis, and Capillary Electrophoresis egg Not available for the exactly traditional Western analysis of the advantage of white immunoassay system.So, optimal system should be in conjunction with two The advantage of person, the shortcoming simultaneously overcoming both.

Content of the invention

The technical problem to be solved is, the shortcoming overcoming prior art, provides one kind to be capable of automatically The protein chromatographic electrophoresis of change process and its in-situ chemical trace and immune imaging device and method.

In order to solve above technical problem, the present invention provides protein chromatographic electrophoresis and its in-situ chemical trace to become with immunity As device, including multichannel chromatoelectrophoresis module, the first solution tank, the multichannel chromatoelectrophoresis of multiple tracks is adopted to separate Plate, active chromatography media and LED fluorescence light source, the liquid-phase outlet of described multichannel chromatoelectrophoresis module is connected to multichannel chromatography electricity One end of swimming separating plate, the another of multichannel chromatoelectrophoresis separating plate is connected to the first solution tank, and described activity chromatography media is put In the track of chromatoelectrophoresis separation flat board, described LED fluorescence light source is used for irradiating described multichannel chromatoelectrophoresis separation Plate, described activity chromatography media includes particulate, and described particulate is surface active ion exchange resin, affinity chromatography filler, anti- Phase chromatographic stuffing or nonporous silica silicon particle or cellular glass particulate or macromolecular material particulate, in described particulate Surface is covalently bonded with two a word used for translations and suckes class coupling molecule.

Being further defined to of technical solution of the present invention, described multichannel chromatoelectrophoresis module is cut for multichannel chromatoelectrophoresis Die change block, described solution tank is multichannel chromatoelectrophoresis handover module, and described multichannel chromatoelectrophoresis handover module includes respectively Electrophoresis tank, multi-channel loading hole, multi-channel switch and multiple chromatography liquid channel, have positive electrode respectively and bear in described electrophoresis tank Electrode, multi-channel loading hole is connected with electrophoresis tank, and multi-channel loading hole is connected with track respectively through multi-channel switch, Described chromatography liquid channel is connected with track respectively.

Being further defined to of technical solution of the present invention, described multi-channel switch is multichannel jettron, described One pneumatic switch slot is arranged at the bottom of each well, has a jettron film in each pneumatic switch slot described, will be pneumatic Switch groove is separated into two passages, and described each well passage through pneumatic switch slot is connected with chromatography liquid channel Logical, another passage is connected with jettron gas access, when jettron film inputs malleation from jettron gas access Or during negative pressure driving, jettron film is separately turned on and closes the path between each well and each chromatography liquid channel.

Being further defined to of technical solution of the present invention, also includes micro liquid phase transfer tube, described micro liquid phase transfer tube It is placed in porch and the exit of chromatography liquid channel.

Being further defined to of technical solution of the present invention, described particulate is microballoon that diameter is 10 microns to 100 microns.

Being further defined to of technical solution of the present invention, described multichannel chromatoelectrophoresis separating plate contains multiple 4- by two pieces 12 mm wides, the glass of 0.1-1 millimeter deep trouth, lucite or transparent polymer material plate face bonded face form, and are formed many Individual 4-12 mm wide, the multichannel chromatoelectrophoresis separating tank of 0.2-2 millimeters deep, two ends have fixing filler to seal swimming lane mouth, shape Become multiple tracks.

Based on above technical problem, the present invention also provides protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging Method, comprises the steps：1) carry out chromatoelectrophoresis using the active chromatography media in multichannel chromatoelectrophoresis separating plate to separate, Described activity chromatography media includes particulate, and described particulate is surface active ion exchange resin, affinity chromatography filler, inversion layer Analysis filler or nonporous silica silicon particle or cellular glass particulate or macromolecular material particulate, 2) in described active layer In analysis medium, particulate is the particulate that its surface is covalently bonded with that two a word used for translations suck class coupling molecule, with micro- described in the light irradiation of specific wavelength Grain, two a word used for translations activating described chromatoelectrophoresis dielectric surface suck quasi-molecule, make two a word used for translations of activation suck the protein of quasi-molecule and surrounding There is chemical coupling, be covalently bound to described microparticle surfaces, make protein produce in-situ chemical trace in described microparticle surfaces；3) First antibody is transported to mutual with the in-situ chemical trace protein that chemical coupling occurs in multichannel chromatoelectrophoresis separating plate Effect；4) wash unreacted first antibody off, by SA be transported in multichannel chromatoelectrophoresis separating plate with first antibody Interact；When SA is fluorescently-labeled, irradiate the surface of SA with LED fluorescence light source, excite fluorescence labeling Luminous, directly observe or by the camera with respective filter directly over multichannel chromatoelectrophoresis separating plate or take the photograph As camera lens is observed and records the surging image of protein, complete protein fluorescence immune imaging；If using enzyme connection mark SA, by enzyme connection mark luminous substrate be transported in multichannel chromatoelectrophoresis separating tank with enzyme join label phase interaction With and light, directly observe or observed by the camera directly over multichannel chromatoelectrophoresis separating plate or pick-up lens and The surging image of record protein, completes the imaging of protein chemistry electrochemiluminescent immunoassay.

Being further defined to of technical solution of the present invention, described step 1 adopts protein chromatographic as claimed in claim 2 Electrophoresis and its in-situ chemical trace and immune imaging device, comprise the following steps that：1a) close multi-channel switch, described leading to made more Path blockade between road well and described track, by described chromatography liquid channel to described multichannel chromatoelectrophoresis Chromatoelectrophoresis buffer solution required for adding in separating tank, adding in described protein well needs the albumen of chromatographic analysis Quality sample, adds electrophoretic buffer, 1b in electrophoresis tank) open multi-channel switch, make described multi-channel loading hole and described electricity Path between swimming swimming lane is opened, and adds suitable voltage simultaneously, make on left and right two described multichannel chromatoelectrophoresis handover modules Protein example in described protein well moves in the electric field in described multichannel chromatoelectrophoresis separating tank and is divided From.

Being further defined to of technical solution of the present invention, described step 1 adopts protein chromatographic as claimed in claim 2 Electrophoresis and its in-situ chemical trace and immune imaging device, comprise the following steps that：1a) close multi-channel switch, described leading to made more Path blockade between road well and described track, by described chromatography liquid channel to described multichannel chromatoelectrophoresis Chromatoelectrophoresis buffer solution required for adding in separating tank, adding in described protein well needs the albumen of chromatographic analysis Quality sample, adds electrophoretic buffer, 1b in electrophoresis tank) open multi-channel switch, make described multi-channel loading hole and described electricity Path between swimming swimming lane is opened, and adds suitable voltage simultaneously, make on left and right two described multichannel chromatoelectrophoresis handover modules Protein example in described protein well moves to the start bit of described multichannel chromatoelectrophoresis separating tank in the electric field Put, 1c) close multi-channel switch, make the path blockade between described multi-channel loading hole and described track；Meanwhile, pass through Required chromatographic flow phase is inputted multichannel chromatoelectrophoresis separating tank by described chromatography liquid channel, makes protein example in described layer Separated in analysis electrophoretic medium.

Being further defined to of technical solution of the present invention, described step 4) comprise the following steps that：4) close multi-channel switch, Make the path blockade between described multi-channel loading hole and described track, to described led to by described chromatography liquid channel more Road chromatography electrophoretic separation groove in sequentially input buffer solution, one or more be capable of identify that the first antibody of some protein, fluorescence Or the SA of enzyme connection mark.

Based on above technical problem, the present invention also provides multifunctional protein chromatoelectrophoresis and its recovery method, according to power Profit requires the protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging device described in 2, comprises the steps：1a) close Close multi-channel switch, make the path blockade between described multi-channel loading hole and described track, by described chromatography liquid phase Passage adds required chromatoelectrophoresis buffer solution in described multichannel chromatoelectrophoresis separating tank, in described protein well Middle addition needs the protein example of chromatographic analysis, adds electrophoretic buffer, 1b in electrophoresis tank) open multi-channel switch, make Path between described multi-channel loading hole and described track is opened, simultaneously in left and right two described multichannel chromatography electricity Add suitable voltage on swimming handover module, make the protein example in described protein well moves to described leading in the electric field more Separated in road chromatography electrophoretic separation groove；1c) close multi-channel switch, make described multi-channel loading hole and described track Between path blockade；Meanwhile, by described chromatography liquid channel, required chromatographic flow phase input multichannel chromatoelectrophoresis is divided From groove, the protein example after making to separate sequentially is exported simultaneously by the liquid-phase outlet of right multichannel chromatoelectrophoresis handover module Collected respectively with collector.

Based on above technical problem, the present invention also provides multifunctional protein chromatoelectrophoresis method, according to claim 3 institute The protein chromatographic electrophoresis stated and its in-situ chemical trace and immune imaging device, comprise the steps：1a) adjust described pneumatic Gas pressure in switch groove, makes described jettron film be under barotropic state, closes described multi-channel loading hole and described layer Path between analysis liquid channel, is added required in described multichannel chromatoelectrophoresis separating tank by described chromatography liquid channel The chromatoelectrophoresis buffer solution wanted, adding in described protein well needs the protein example of chromatographic analysis, in electrophoresis tank Interior addition electrophoretic buffer, 1b) adjust gas pressure in described pneumatic switch slot, make described jettron film be in negative pressure state Under, open the path between described multi-channel loading hole and described chromatography liquid channel, simultaneously in left and right two described multichannels Add suitable voltage on chromatoelectrophoresis handover module, make the protein example in described protein well move to institute in the electric field State in multichannel chromatoelectrophoresis separating tank and separated.

The invention has the beneficial effects as follows：One of innovative point of this patent is exactly unique chromatoelectrophoresis medium and is chromatographed with this Electrophoretic medium replaces polyacrylamide gel, on the one hand makes protein can complete to separate in chromatoelectrophoresis medium, so that it is had There is the preparation of traditional protein electrophoresis and commonly use, further aspect is that its surface is wrapped in one layer of chemical coupling Material, can activate under the LED light induction of special wavelength, in the original location coupling reaction occur with albumen, make albumen covalent bond To on chromatoelectrophoresis medium.Complete the original position solid printing process of traditional protein electrophoresis.These innovative points are all Capillary Electrophoresis Cannot complete, be a novelty transformation to the analysis of Capillary Electrophoresis related immune.These easily conditions, convenient realize The complete of WESTERN instrument is changed certainly.The present invention has been effectively combined traditional LC electrophoresis, chemical quick coupling characteristic, tradition Western analyzes the advantage of three kinds of technology, have developed a kind of new chromatoelectrophoresis (electrochromatography, electro Chromatography, the electrophoresis that finger protein is carried out in LC medium, at present only in Capillary Electrophoresis using), can So that conventional Western is analysis automated, thus setting up full-automatic Western instrument.Firstly, since full-automatic Western instrument is There are preparation and the imaging advantage of traditional Western, have the automation of Capillary Electrophoresis and quick analytic function, it is than tradition again Western analysis there is more practical and wider using value.Full-automatic Western instrument drastically increases The speed of Western analysis, that is, time-consuming, save manpower again.In addition, it is not necessary that senior technician, can reduce artificial Operating error, be easier promote, reduce the use threshold of Western technology.The biological peace of full-automatic Western instrument Quan Xing：Due to cannot be avoided the contact of operating personnel and albumen sample in traditional Western analysis, especially it is infectious in analysis During the clinical samples of disease, operating personnel are subject to potential threat unavoidably.The use of full-automatic Western instrument can be to greatest extent Protect operating personnel from the threat having potential infectiousness sample, eliminate biological safety hidden danger.Can be using full-automatic Western instrument includes 1 come the pathogen to detect) virus：HIV, HCV, HEV, cytomegalovirus, herpes simplex virus, tiny disease Poison, rubella virus, Epstein-Barr virus etc.；2) bacterium：Conveyor screw, Brucella, helicobacter pylori, syphilis, Yersinia；3) post Infested：Toxoplasmosis etc..

The significance of full-automatic Western instrument：Full-automatic Western instrument can improve life science and clinical doctor Learn the ability of diagnosis, at aspects such as life science, clinical medicine diagnosis and disease detection controls, all there is huge application Prospect.In terms of clinical diagnosis, the appearance of full-automatic Western instrument will make Western analytical technology widely be adopted. In terms of life science, because Western is to confirm that protein expression, molecular weight, isoelectric point and protein-interacting etc. must Indispensable standard method, the use of full-automatic Western instrument will be greatly enhanced the speed published an article and quality.Automatically Western instrument is applied to university, medical research unit, hospital laboratory, Center for Disease Control, inspection and quarantine system, center in fact Test room etc..So, the development of full-automatic Western instrument has very high practicality, and the development to modern life science has Significance.

Brief description

Fig. 1 is present protein chromatoelectrophoresis and its in-situ chemical trace and immune imaging device cross-sectional schematic；

Fig. 2 is the chromatoelectrophoresis switching of present protein chromatoelectrophoresis and its in-situ chemical trace and immune imaging device Module cut-away illustration；

Fig. 3 is present protein chromatoelectrophoresis and its in-situ chemical trace separates with the chromatoelectrophoresis of immune imaging device Plate schematic perspective view；

In figure, 1 chromatoelectrophoresis handover module, 2 chromatoelectrophoresis separating plates, 3 chromatoelectrophoresis media, 4LED fluorescence light source, 5 is micro- Amount liquid phase transfer tube, 6 iontophoretic electrodes, 7 electrophoresis tanks, 8 wells, 9 jettron air intakes, 10 jettron films, 11 electrophoresis Swimming lane.

Specific embodiment

Embodiment one

A kind of achievable protein chromatographic electrophoresis and its in-situ chemical trace medium, including chromatography media, described chromatography is situated between Matter is ion exchange resin, various affinity chromatography medium and the reversed phase chromatography microballoon of 10-100 micron diameter, described chromatography media It is filled in the described track 11 of multichannel chromatoelectrophoresis separating plate 2, when being filled with difference in described track 11 Chromatography media when, add chromatography liquid phase (mobile phase), protein will according to distribution of charges and hydrophobicity etc. carry out separate.Micro- Ball surface is covalently bonded with two a word used for translations and suckes class coupling molecule.Described microballoon is coupled through sucking class compound with two a word used for translations, forms uniqueness Two a word used for translations suck compound parcel microballoon, when microballoon described in the light irradiation with specific wavelength, activate two a word used for translations of described chromatographic medium surface Suck quasi-molecule, make two a word used for translations of activation suck quasi-molecule, with the protein of surrounding, chemical coupling occurs, be covalently bound to described microballoon table Face, makes protein produce in-situ chemical trace in described microsphere surface.

Embodiment two

A kind of protein chromatographic electrophoresis and its in-situ chemical trace medium, including electrophoretic medium, described electrophoretic medium includes Particulate, a diameter of 20-50 micron non-porous silicas microballoon of described particulate or cellular glass microballoon or other macromolecular materials Microballoon.Described microballoon is coupled through sucking class compound with two a word used for translations, forms two unique a word used for translations and suckes compound parcel microballoon, with described two A word used for translation suckes compound parcel microballoon as chromatoelectrophoresis medium 3 replacement conventional polypropylene acrylamide gel, can carry out electricity to protein Swimming lock out operation.It is filled in described multichannel with silicon dioxide microsphere or cellular glass microballoon or other macromolecular material microballoon In the track 11 of chromatoelectrophoresis separating plate 2, in addition, work as being filled with polysaccharide or ampholytes etc. in described track 11 During medium, protein is acted on by electrophoretic buffer composition and silicon dioxide microsphere surface molecular in electrophoretic medium, makes not Press its molecular characterization, the such as characteristic such as molecular weight and isoelectric point with swimming in described chromatoelectrophoresis medium 3 for the protein, reach layer The purpose of analysis electrophoretic separation.

Described microballoon is coupled through sucking class compound with two a word used for translations, forms two unique a word used for translations and suckes compound parcel microballoon, when with Particulate described in the light irradiation of specific wavelength, two a word used for translations activating described chromatographic medium surface suck quasi-molecule, make two a word used for translations of activation suck class There is chemical coupling with the protein of surrounding in molecule, be covalently bound to described microparticle surfaces, make protein in described microparticle surfaces Produce in-situ chemical trace.

Embodiment three

As shown in Figure 1 to Figure 2, a kind of protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging device, are commonly called as Full-automatic Western instrument, it includes multichannel chromatoelectrophoresis module, multichannel chromatoelectrophoresis separating plate 2, is placed in multichannel chromatography The LED fluorescence light source 4 of underface of electrophoretic separation plate 2 and be placed in photograph machine machine directly over multichannel chromatoelectrophoresis separating plate 2 or Pick-up lens.Multichannel chromatoelectrophoresis module is responsible for for protein, electrophoretic buffer and chromatographic solution etc. being transported to multichannel chromatography Carry out separating treatment, described multichannel chromatoelectrophoresis separating plate 2 receiving carries out detached electrophoresis chromatography and is situated between in electrophoretic separation plate 2 Matter.The list that described LED fluorescence light source 4 can provide activation photochemical reaction and excite wavelength required for FLA colour developing Color light source.Wherein, wherein LED fluorescence light source 4 forms spot light matrix by the LED of multiple specific wavelengths it is ensured that described chromatography Two a word used for translations on electrophoretic medium 3 surface are suck quasi-molecule molecule and are fully activated.Described photograph or pick-up lens are used for observing and record albumen Matter surging image.Include particulate in described electrophoresis chromatography media, be covalently bonded with two a word used for translations on the surface of described particulate and suck class occasionally Connection molecule.The microballoon of a diameter of 10 microns to 100 microns of described particulate.Described particulate can be ion exchange resin, affine Chromatography media, reversed phase chromatographic medium particulate or nonporous silica silicon particle or cellular glass particulate or other macromolecular material are micro- Ball.Multichannel chromatoelectrophoresis module includes two chromatoelectrophoresis handover modules 1, respectively negative pole chromatoelectrophoresis handover module 1 He Positive pole chromatoelectrophoresis handover module 1, negative pole chromatoelectrophoresis handover module 1 and positive pole chromatoelectrophoresis handover module 1 are respectively placed in electricity The left and right sides of swimming swimming lane 11.Chromatoelectrophoresis handover module 1 and described multichannel chromatoelectrophoresis separating plate 2 on a plane, its In at least one chromatoelectrophoresis handover module 1 can slidably reciprocate in plane and realize chromatoelectrophoresis handover module 1 and electrophoresis The separation of swimming lane 11 merges；When two chromatoelectrophoresis handover modules 1 all separate, multichannel chromatoelectrophoresis separating plate 2 can be certainly By taking out or put back to；When two chromatoelectrophoresis handover modules 1 merge, multiple outlets of chromatoelectrophoresis handover module 1 respectively with The track 11 of multichannel chromatoelectrophoresis separating plate 2 aligns；Chromatoelectrophoresis handover module 1 and multichannel chromatoelectrophoresis separating plate 2 binding sites have rubber ring or film, unimpeded but tightly water-tight gentle with the external world to guarantee track 11.As shown in figure 3, it is many Passage chromatography electrophoretic separation plate 2 is multiple tracks 11.Multigroup sample just can be checked so when carrying out sample survey simultaneously Product, described track 11 is upper and lower two blocks of glass with cutting or poly (methyl methacrylate) plate fastens and forms, and specifically, adopts two Block is carved with 0.1-1 millimeters deep, 6-12 mm wide, the glass of the long cutting of 60-120 millimeter or transparent acrylic material, glues face-to-face Form 0.2-2 millimeters thick, 6-12 mm wide, what 60-120 millimeter was long can accommodate the track of chromatography electrophoretic medium 3 after conjunction 11, two ends have fixing filler to seal swimming lane mouth, allow to prevent particulate from flowing out while liquid phase flowing.Described fixing filler is permissible It is molded in the following way：Add a small amount of 20 micron silica microballoons and formyl in chromatoelectrophoresis swimming lane 11 one end being formed Amine simultaneously makes it be coupled solidification at high temperature, forms the silicon dioxide microsphere cured layer of about 2-5 millimeters thick.Described silica Microballoon cured layer will allow for liquid to be passed through, but can stop the outflow of described chromatoelectrophoresis medium 3.Ion equally can also be adopted Exchanger resin or reversed phase chromatography microballoon and formamide simultaneously make it be coupled solidification at high temperature, formed about 2-5 millimeters thick from Sub-exchange resin or reversed phase chromatography microballoon cured layer.As shown in Fig. 2 described multichannel chromatoelectrophoresis handover module 1 wraps respectively Include electrophoresis tank 7, multi-channel loading hole 8, multichannel jettron and multiple chromatography liquid channel, have respectively in described electrophoresis tank 7 Positive electrode and negative electrode, multi-channel loading hole 8 is connected with electrophoresis tank 7, and multi-channel loading hole 8 is through multichannel jettron respectively It is connected with chromatography liquid channel, described chromatography liquid channel is connected with track 11 respectively.The plurality of chromatography liquid phase The entrance of passage or exit are equipped with micro liquid phase transfer tube 5.There is a jettron in the bottom of each well 8 described Groove, has a jettron film 10 in each pneumatic switch slot described, pneumatic switch slot is separated into two passages, and described is every A passage through pneumatic switch slot for the individual well 8 is connected with chromatography liquid channel, another passage and jettron air Entrance 9 is connected, and jettron air intake 9 is connected with inlet valve and air outlet valve, when jettron film 10 is from jettron air When entrance 9 input malleation or negative pressure drive, jettron film 10 is separately turned on and closes each well 8 chromatographing liquid phase with each Path between passage.For carrying out electrophoresis and chromatography.

For foregoing invention, its technical characterstic is to carry out protein chromatographic and the two kinds of runnings of performance liquid electrophoresis Pattern.And described chromatoelectrophoresis modular converter can be cut between protein chromatographic and two kinds of operating modes of protein electrophorese Change, reach the purpose completing protein chromatographic and two kinds of operations of protein electrophorese.For above-mentioned chromatoelectrophoresis handover module 1, when When being in electrophoretic, the jettron air intake 9 of described chromatoelectrophoresis handover module 1 is in negative pressure, jettron Film 10 is in opening, and positive and negative iontophoretic electrode 6 can pass through electrophoretic buffer, via well 8 and jettron film 10 with The track 11 of multichannel chromatoelectrophoresis separating plate 2 is connected, and forms electrophoresis loop.(or:By negative pole chromatoelectrophoresis handover module 1 electrophoresis tank 7 cataphoresis electrode 6 enters well 8 and enters jettron film 10, enters multichannel chromatoelectrophoresis separating plate 2 Track 11, enters back into the positive pole of the electrophoresis tank 7 of positive pole chromatoelectrophoresis handover module 1.) when being in chromatography pattern, described The jettron air intake 9 of chromatoelectrophoresis handover module 1 be generally in normal pressure, jettron film 10 is in closing shape State, now jettron film 10 turn off, multichannel chromatoelectrophoresis separating plate 2 and positive and negative iontophoretic electrode 6 and electrophoretic buffer are by gas Dynamic switch membrane 10 separates, and electrophoresis loop is interrupted, the electrophoresis of micro liquid phase transfer tube 5 and multichannel chromatoelectrophoresis separating plate 2 simultaneously Swimming lane 11 is connected, and forms LC loop.The solution of needs and reagent can be transported to by described micro liquid phase transfer tube 5 In track 11.

When opening multi-channel switch, the path between described multi-channel loading hole 8 and described track 11 is made to open, this When under electrophoretic, in the presence of electric field protein can from chromatoelectrophoresis handover module 1 enter multichannel chromatoelectrophoresis Separated in the track 11 of separating plate 2 and in chromatoelectrophoresis medium 3.When closing multi-channel switch, made described leading to more Path blockade between road well 8 and described track 11, now under chromatography pattern, solution and protein are in micro liquid Multichannel chromatoelectrophoresis separating plate 2 is entered and in chromatoelectrophoresis medium from chromatoelectrophoresis handover module 1 under the promotion of phase transfer tube 5 In 3, chromatography is carried out it is possible to enter line replacement to liquid phase part to albumen.It should be noted that：Under chromatography pattern, sample-adding Hole 8 and track 11 are not communicated with, in order that protein can enter in track 11 by jettron film 10, then Jettron film 10 should frequently be opened, simultaneously in the micro liquid phase transfer tube 5 of negative pole or positive pole chromatoelectrophoresis handover module 1 Under driving, protein is swum to positive pole from negative pole.In order to realize the chromatography of protein, can also adopt alternatively：I.e. in advance By electrophoretic, protein is transported to the original position of multichannel chromatoelectrophoresis separating tank, comprises the concrete steps that：Adjust described Gas pressure in pneumatic switch slot, makes described jettron film 10 be under negative pressure state, open described multi-channel loading hole 8 with Path between described chromatography liquid channel, on left and right two described multichannel chromatoelectrophoresis handover modules 1, adduction is fitted simultaneously Voltage, makes the protein example in described protein well 8 move to described multichannel chromatoelectrophoresis separating tank in the electric field Original position.When for chromatography mode of operation, will be with generation in-situ chemical trace egg under the driving of micro liquid phase transfer tube 5 White matter occurs the first antibody of immune imaging and the luminous substrate of SA and enzyme connection mark to send into multichannel chromatoelectrophoresis and divide In plate 2.

For above-described embodiment, it can also be multiple that electrophoresis tank 7 can be one, and in addition it can also individually be swum with electrophoresis Road 11 is connected, as long as protein is just permissible through the conductive path of electrophoresis tank 7.It can set on the path of electrophoresis tank 7 Put accumulator tank, so that the output end of protein well 8 is connected with accumulator tank, you can ensure completing of electrophoresis process.Described LED Fluorescence light source 4 can also be placed in the top of multichannel chromatoelectrophoresis separating plate 2, as long as or other positions can be irradiated to separate Particulate in pond.

The multichannel chromatoelectrophoresis separating plate 2 of above-mentioned track 11 is that one kind can use described chromatoelectrophoresis medium 3 electrophoretic apparatus, its effect is to allow described chromatoelectrophoresis medium 3 be formed as the physical size of conventional polypropylene acrylamide gel With shape it is ensured that protein can be effectively separated under chromatography and electrophoresis both of which, guarantee that protein can be simultaneously The operation such as in-situ chemical trace and immune imaging is completed in chromatoelectrophoresis medium 3.

Present invention additionally comprises a kind of chromatoelectrophoresis medium 3 for replacing conventional polypropylene acrylamide gel.Described chromatography electricity The technical requirements of swimming medium 3 include 1) replace conventional polypropylene acrylamide gel but still protein electrophorese separation can be carried out, 2) Allow 1-2000 mul/min of liquid flow velocity it is ensured that the flowing in chromatoelectrophoresis medium 3 of buffer solution and solution, 3) There is the chemical reaction characteristic of coupling protein matter, under the activation of light or chemical reagent, with peripheral protein matter, chemistry occurs Coupling reaction.

Example IV

It is with embodiment three difference, chromatoelectrophoresis handover module 1 only one of which, it is located at multichannel chromatoelectrophoresis The left side of separating plate 2, right side only has the first solution tank, has positive electrode in described first solution tank.First solution tank passes through layer Analysis liquid channel is connected with track 11, has negative electrode, in addition in the electrophoresis tank 7 of described chromatoelectrophoresis handover module 1 Described micro liquid phase transfer tube 5 is one, and it is located at entrance or the outlet of chromatography liquid channel.

Embodiment five

A kind of protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging device, including multichannel chromatoelectrophoresis mould Block, the first solution tank, the multichannel chromatoelectrophoresis separating plate 2 using multiple tracks 11, active chromatography media and LED fluorescence Light source 4, the liquid-phase outlet of described multichannel chromatoelectrophoresis module is connected to one end of multichannel chromatoelectrophoresis separating plate 2, multichannel The another of chromatoelectrophoresis separating plate 2 is connected to the first solution tank, and described activity chromatography media is placed in chromatoelectrophoresis separation flat board In track 11, described LED fluorescence light source 4 is used for irradiating described multichannel chromatoelectrophoresis separating plate 2.Described multichannel chromatography Electrophoresis module is multichannel chromatoelectrophoresis handover module 1, and described multichannel chromatoelectrophoresis handover module 1 includes the second solution respectively Groove and multi-channel loading hole 8, described second solution tank is connected with track 11, and described multi-channel loading hole 8 and electrophoresis are swum Road 11 is connected, and has positive electrode and negative electrode respectively in described first, second solution tank.This structure can also be realized chromatographing mould Formula and electrophoretic, under chromatography pattern, the luminous substrate of solution, first antibody and SA and enzyme connection mark is molten from second Liquid bath can also be from multichannel chromatoelectrophoresis separating plate 2 while shift to another side by gravity.

For above-described embodiment, in fact chromatoelectrophoresis module can adopt various structure, such as using independent layer Analyse conveyor module or using independent electrophoresis conveyor module, well 8 can adopt individually independent structure and track 11 are connected, and first, second solution tank is then connected with track 11 by another passage.Well 8 can also be not only Vertical exist, but directly carries out protein sample-adding by solution tank.And these are simply simply enumerated, in fact also have many kinds Other structures realized, these structures should not depart from protection scope of the present invention.

Above-mentioned independent chromatography conveyor module can adopt following structure：Described multichannel chromatoelectrophoresis module includes molten Liquid bath and multi-channel loading hole 8, described solution tank is connected with track 11, described multi-channel loading hole 8 and track 11 are connected.This multichannel chromatoelectrophoresis module is two, the one end being located at multichannel chromatoelectrophoresis separating plate 2, another The individual other end positioned at multichannel chromatoelectrophoresis separating plate 2.

Above-mentioned independent electrophoresis conveyor module can adopt following structure：Described multichannel chromatoelectrophoresis module includes molten Liquid bath and multi-channel loading hole 8, described solution tank is connected with well 8, described multi-channel loading hole 8 and track 11 phase Connection.This multichannel chromatoelectrophoresis module is two, the one end being located at multichannel chromatoelectrophoresis separating plate 2, another position The other end in multichannel chromatoelectrophoresis separating plate 2.Solution positioned at one end of multichannel chromatoelectrophoresis separating plate 2 and the other end There are negative electrode and positive electrode respectively in groove.

Embodiment six

It is with embodiment three difference：Described multichannel chromatoelectrophoresis module is multichannel chromatoelectrophoresis handover module 1, described multichannel chromatoelectrophoresis handover module 1 includes electrophoresis tank 7, multi-channel loading hole 8, multi-channel switch and multiple layer respectively Analysis liquid channel, has positive electrode or negative electrode, multi-channel loading hole 8 is connected with electrophoresis tank 7, many respectively in described electrophoresis tank 7 Passage well 8 is connected with track 11 respectively through multi-channel switch, described chromatography liquid channel respectively with track 11 are connected.Described multi-channel switch can be various forms of switches, such as mechanical switch etc..

Embodiment seven

A kind of protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging method, comprise the steps：1) utilize Active chromatography media in multichannel chromatoelectrophoresis separating plate 2 carries out electrophoresis chromatography, and described activity chromatography media includes Particulate, described particulate is surface active ion exchange resin, affinity chromatography filler, reversed phase chromatography filler or non-porous silicas Particulate or cellular glass particulate or macromolecular material particulate, when for nonporous silica silicon particle or cellular glass particulate or During person's macromolecular material particulate, described step 1 is using the protein chromatographic electrophoresis as described in embodiment six and its in-situ chemical print Mark and immune imaging device, described step 1) comprise the following steps that：1a) close multi-channel switch, make described multi-channel loading Path blockade between hole 8 and described track 11, is divided to described multichannel chromatoelectrophoresis by described chromatography liquid channel Chromatoelectrophoresis buffer solution required for adding in groove, adding in described protein well 8 needs the albumen of chromatographic analysis Quality sample, in electrophoresis tank 7 add electrophoretic buffer, 1b) open multi-channel switch, make described multi-channel loading hole 8 with described Path between track 11 is opened, and adds appropriate electrical on left and right two described multichannel chromatoelectrophoresis handover modules 1 simultaneously Pressure, makes the protein example in described protein well 8 move in the electric field in described multichannel chromatoelectrophoresis separating tank Separated.2) in described active chromatography media, particulate is that its surface is covalently bonded with two a word used for translations and suckes the micro- of class coupling molecule Grain, with particulate described in the light irradiation of specific wavelength, two a word used for translations activating described chromatoelectrophoresis medium 3 surface suck quasi-molecule, make activation Two a word used for translations suck the protein of quasi-molecule and surrounding chemical coupling occur, be covalently bound to described microparticle surfaces, make protein in institute State microparticle surfaces and produce in-situ chemical trace.3) after completing in-situ chemical trace, chromatoelectrophoresis handover module 1 is switched to First antibody is transported to chemical with generation in multichannel chromatoelectrophoresis separating plate 2 by chromatography pattern by micro liquid phase transfer tube 5 The in-situ chemical trace protein interaction being coupled；4) wash unreacted first antibody off, will by micro liquid phase transfer tube 5 SA is transported in multichannel chromatoelectrophoresis separating plate 2 and is interacted with first antibody；When SA is fluorescence labeling , irradiate the surface of SA with LED fluorescence light source 4, excite fluorescence labeling to light, directly observe or by positioned at multichannel The camera with respective filter directly over chromatoelectrophoresis separating plate 2 or pick-up lens are observed and are recorded springing up of protein Image, completes protein fluorescence immune imaging.If using the SA of enzyme connection mark, such as SA is peroxidating Enzyme mark, by micro liquid phase transfer tube 5, the luminous substrate of peroxidase is transported in multichannel chromatoelectrophoresis separating tank Interact with peroxidase and light, directly observe or by the photograph directly over multichannel chromatoelectrophoresis separating plate 2 Machine or pick-up lens are observed and are recorded the surging image of protein, complete the imaging of protein chemistry electrochemiluminescent immunoassay.Described step 4) Comprise the following steps that：4) close multi-channel switch, so that the path between described multi-channel loading hole 8 and described track 11 is closed Close, by described chromatography liquid channel sequentially input in described multichannel chromatoelectrophoresis separating tank buffer solution, one or more It is capable of identify that the SA of the first antibody, fluorescence or enzyme connection mark of some protein.

Present invention also offers a kind of side that fluorescent marker protein in described chromatoelectrophoresis medium 3 is carried out with real-time monitoring Method.When chromatoelectrophoresis being carried out to fluorescent marker protein, by opening LED fluorescence light source 4, excite in chromatography track 11 Fluorescent marker protein, by naked eyes or camera installation, springs up to protein and carries out Real Time Observation and imaging.

When being separated by electrophoresis using ampholytes or polysaccharide, the method for operating of the present embodiment is as follows：In two layers Add appropriate electrophoretic buffer in the electrophoresis tank 7 of analysis electrophoresis handover module 1, electrophoretic buffer is used by micro liquid phase transfer tube 5 Fluid pipeline in chromatoelectrophoresis handover module 1 and multichannel chromatoelectrophoresis separating plate 2 is rinsed, is driven by micro liquid phase Dynamic pump 5 is with being balanced to multichannel chromatoelectrophoresis separating plate 2 containing ampholytes or polysaccharide.

When filling the track 11 of multichannel chromatoelectrophoresis separating plate 2 using ampholytes (ampholyte), egg White matter can be carried out separating (isoelectric focusing) by isoelectric point；If now not containing silicon dioxide microsphere in chromatoelectrophoresis medium 3, Cannot be carried out in-situ chemical trace and immune imaging.

When filling the track 11 of multichannel chromatoelectrophoresis separating plate 2 using the electrophoretic buffer containing polysaccharide, egg White matter can be carried out separating by molecular size range.If now not containing silicon dioxide microsphere in chromatoelectrophoresis medium 3, cannot enter Row in-situ chemical trace and immune imaging.

Protein example is added in the well 8 of chromatoelectrophoresis handover module 1, by chromatoelectrophoresis handover module 1 It is switched to electrophoretic and starts electrophoresis, protein will enter multichannel chromatoelectrophoresis from well 8 and divide in the presence of electric field Separated from plate 2 and in chromatoelectrophoresis medium 3.

Above-mentioned protein example can also change fluorescently-labeled protein into.

After bromophenol blue indicator goes to suitable position in multichannel chromatoelectrophoresis separating plate 2, open described LED Fluorescence light source 4, two a word used for translations on activation chromatography electrophoretic medium 3 surface suck class coupling molecule, and induction is even with the free radical of peripheral protein matter Connection reaction, completes the in-situ chemical trace of protein surging image.

Chromatography pattern is switched to by chromatoelectrophoresis handover module 1, is driven by micro liquid phase by micro liquid phase transfer tube 5 First antibody is transported to the protein interaction with coupling in chromatoelectrophoresis medium 3 by the input port of pump 5.

Under chromatography pattern, unreacted first antibody is washed off by micro liquid phase transfer tube 5, then SA is conveyed Interact with first antibody in chromatoelectrophoresis medium 3.

If SA is fluorescently-labeled, described LED fluorescence light source 4 can be opened, excite fluorescence labeling to light, and Observe and record the surging image of protein by the photograph directly over multichannel chromatoelectrophoresis separating plate 2 or pick-up lens, complete Become protein chromatographic electrophoresis and its in-situ chemical trace and fluorescence immunoassay imaging.

If SA is peroxidase mark, by micro liquid phase transfer tube 5 by the luminous substrate of peroxidase Be transported in chromatoelectrophoresis medium 3 with peroxidase interact and light, by camera installation observing protein surging image, Complete protein chromatographic electrophoresis and its in-situ chemical trace and chemiluminescence immunoassay imaging.

When carrying out chromatography using ion exchange or anti-phase affinity chromatography, the method for operating of the present embodiment is as follows：

If using ion exchange or anti-phase affinity chromatography etc., chromatographing electrophoretic medium 3 when replacing using ion exchange resin When, protein will carry out separating according to distribution of charges；Chromatograph electrophoretic medium 3 when replacing using reversed phase chromatographic medium (C8, C18 etc.) When, protein will carry out separating according to hydrophobicity；Protein example is added in the well 8 of chromatoelectrophoresis handover module 1 After be switched to chromatography pattern, by protein and layer under the driving of the micro liquid phase transfer tube 5 of positive pole chromatoelectrophoresis handover module 1 Analysis liquid enters multichannel chromatoelectrophoresis separating plate 2 from well 8 and is separated chromatoelectrophoresis medium 3.Noticeable It is：Under chromatography pattern, well 8 and track 11 are not communicated with, in order that protein can pass through jettron film 10 Enter in track 11, then should frequently open jettron film 10, switch in negative pole or positive pole chromatoelectrophoresis simultaneously Under the micro liquid phase transfer tube 5 of module 1 drives, protein is swum to positive pole from negative pole.In addition can also be beforehand through electrophoretic Protein is transported to the original position of multichannel chromatoelectrophoresis separating tank.

Other steps are with identical when being separated by electrophoresis using ampholytes or polysaccharide.

Above-mentioned protein can also be fluorescently-labeled protein.

Because protein is fluorescently-labeled, described LED fluorescence light source 4 can be opened, excite fluorescence labeling to light, and lead to Photograph directly over excessive passage chromatography electrophoretic separation plate 2 or the surging image of pick-up lens observation and record protein.

Embodiment eight

It is step 1 with embodiment seven difference：Described step 1 comprises the following steps that：1a) close multi-channel switch, Make the path blockade between described multi-channel loading hole 8 and described track 11, by described chromatography liquid channel to described Chromatoelectrophoresis buffer solution required for adding in multichannel chromatoelectrophoresis separating tank, adding in described protein well 8 needs Want the protein example of chromatographic analysis, add electrophoretic buffer, 1b in electrophoresis tank 7) open multi-channel switch, made described leading to more Path between road well 8 and described track 11 is opened, simultaneously in left and right two described multichannel chromatoelectrophoresis switchings Add suitable voltage in module 1, make the protein example in described protein well 8 move to described multichannel layer in the electric field The original position of analysis electrophoretic separation groove, 1c) close multi-channel switch, make described multi-channel loading hole 8 and described track 11 Between path blockade；Meanwhile, by described chromatography liquid channel, required chromatographic flow phase input multichannel chromatoelectrophoresis is divided From groove, protein example is made to be separated in described chromatoelectrophoresis medium 3.

Embodiment nine

The present embodiment provides a kind of novel protein chromatoelectrophoresis and its retracting device, protein can be pressed isoelectric point, Molecular weight or other molecular characterization carry out separating and can reclaim the protein after separating, that is, increased auto partial sampler, Described auto partial sampler is 96- orifice plate part collector.Other parts are identical with embodiment three.Described in the present embodiment from Dynamic part collector is placed in the downstream of positive pole chromatoelectrophoresis handover module 1, and the micro liquid phase of positive pole chromatoelectrophoresis handover module 1 is driven After dynamic pump 5.Multichannel chromatoelectrophoresis is separated by the micro liquid phase transfer tube 5 that can pass through positive pole chromatoelectrophoresis handover module 1 Protein in the track 11 of plate 2 slowly pumps out, and is collected by 96 components by auto partial sampler.Wherein 96- Orifice plate auto partial sampler can be existing auto partial sampler on market.

The method of operating of the present embodiment approximately as：

Performance liquid is separated by electrophoresis：The first step, in the presence of electric field, fluorescent marker protein will enter from well 8 Enter multichannel chromatoelectrophoresis separating plate 2 and separated in chromatoelectrophoresis medium 3.Second step, opens described LED fluorescence light source 4, excite fluorescence labeling to light, and by the photograph directly over multichannel chromatoelectrophoresis separating plate 2 or pick-up lens record albumen The surging image of matter.3rd step, is switched to chromatography pattern, in the micro liquid phase transfer tube 5 of positive pole chromatoelectrophoresis handover module 1 Under driving, the fluorescent marker protein after separating in multichannel chromatoelectrophoresis separating plate 2 is pressed by 96- orifice plate part collector Component is collected respectively.

Protein chromatographic separates：The first step, in the driving of the micro liquid phase transfer tube 5 of positive pole chromatoelectrophoresis handover module 1 Lower protein will enter multichannel chromatoelectrophoresis separating plate 2 from well 8 and be separated chromatoelectrophoresis medium 3.Second Step, opens described LED fluorescence light source 4, excites fluorescence labeling to light, and by multichannel chromatoelectrophoresis separating plate 2 surface Photograph or the surging image of pick-up lens record protein.3rd step, drives in the micro liquid phase of positive pole chromatoelectrophoresis handover module 1 Under the driving of dynamic pump 5, the note protein after separating in multichannel chromatoelectrophoresis separating plate 2 is pressed by 96- orifice plate part collector Component is collected respectively.

In addition to the implementation, the present invention can also have other embodiment.All employing equivalents or equivalent transformation shape The technical scheme becoming, all falls within the protection domain of application claims.

Claims (10)

1. protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging device it is characterised in that：Including multichannel chromatography Electrophoresis module, the first solution tank, the multichannel chromatoelectrophoresis separating plate using multiple tracks, active chromatography media and LED Fluorescence light source, the liquid-phase outlet of described multichannel chromatoelectrophoresis module is connected to one end of multichannel chromatoelectrophoresis separating plate, leads to more The another of road chromatography electrophoretic separation plate is connected to the first solution tank, and described activity chromatography media is placed in chromatoelectrophoresis separation flat board In track, described LED fluorescence light source is used for irradiating described multichannel chromatoelectrophoresis separating plate, described activity chromatography media bag Include particulate, described particulate is surface active ion exchange resin, affinity chromatography filler, reversed phase chromatography filler or atresia dioxy SiClx particulate or cellular glass particulate or macromolecular material particulate, are covalently bonded with two a word used for translations on the surface of described particulate and suck class Coupling molecule；Described multichannel chromatoelectrophoresis module is multichannel chromatoelectrophoresis handover module, and described first solution tank is to lead to more Road chromatoelectrophoresis handover module, described multichannel chromatoelectrophoresis handover module includes electrophoresis tank, multi-channel loading hole respectively, leads to more Road switch and multiple chromatography liquid channels, have positive electrode and negative electrode, multi-channel loading hole and electrophoresis respectively in described electrophoresis tank Groove is connected, and multi-channel loading hole is connected with track respectively through multi-channel switch, described chromatography liquid channel respectively with Track is connected.

2. protein chromatographic electrophoresis according to claim 1 and its in-situ chemical trace and immune imaging device, its feature It is：Described multi-channel switch is multichannel jettron, has a pneumatic switch slot in the bottom of each well, described There is a jettron film in each pneumatic switch slot, pneumatic switch slot is separated into two passages, each well is through pneumatic One passage of switch groove is connected with chromatography liquid channel, and another passage is connected with jettron gas access, works as gas When jettron gas access inputs malleation or negative pressure drives, jettron film is separately turned on and closes each adding dynamic switch membrane Path between sample hole and each chromatography liquid channel.

3. protein chromatographic electrophoresis according to claim 2 and its in-situ chemical trace and immune imaging device, its feature It is：Also include micro liquid phase transfer tube, described micro liquid phase transfer tube is placed in porch and/or the outlet of chromatography liquid channel Place.

4. protein chromatographic electrophoresis according to claim 1 and its in-situ chemical trace and immune imaging device, its feature It is：Described particulate is microballoon that diameter is 10 microns to 100 microns.

5. protein chromatographic electrophoresis according to claim 1 and its in-situ chemical trace and immune imaging device, its feature It is：Described multichannel chromatoelectrophoresis separating plate is contained multiple 4-12 mm wides, the glass of 0.1-1 millimeter deep trouth, is had by two pieces Machine glass or transparent polymer material plate face bonded face form, and form multiple 4-12 mm wides, the multichannel of 0.2-2 millimeters deep Chromatoelectrophoresis separating tank, two ends have fixing filler to seal swimming lane mouth, form multiple tracks.

6. protein chromatographic electrophoresis and its in-situ chemical trace and immune imaging method, comprise the steps：1）Using multichannel Active chromatography media in chromatoelectrophoresis separating plate carries out chromatoelectrophoresis and separates, and described activity chromatography media includes particulate, institute State particulate be surface active ion exchange resin, affinity chromatography filler, reversed phase chromatography filler or nonporous silica silicon particle or Person's cellular glass particulate or macromolecular material particulate, 2）In described active chromatography media, particulate is its surface covalent bond There are two a word used for translations to suck the particulate of class coupling molecule, with particulate described in the light irradiation of specific wavelength, activate described chromatoelectrophoresis dielectric surface Two a word used for translations suck quasi-molecule, make the protein that two a word used for translations of activation suck quasi-molecule and surrounding that chemical coupling to occur, be covalently bound to described Microparticle surfaces, make protein produce in-situ chemical trace in described microparticle surfaces；3）First antibody is transported to multichannel chromatography With the in-situ chemical trace protein interaction that chemical coupling occurs in electrophoretic separation plate；4）Wash unreacted first off to resist Body, SA is transported in multichannel chromatoelectrophoresis separating plate and is interacted with first antibody；When SA is fluorescence Mark, irradiate the surface of SA with LED fluorescence light source, excite fluorescence labeling to light, directly observe or many by being located at The camera with respective filter directly over passage chromatography electrophoretic separation plate or pick-up lens are observed and are recorded protein Surging image, completes protein fluorescence immune imaging；If using the SA of enzyme connection mark, by sending out of enzyme connection mark Light substrate is transported in multichannel chromatoelectrophoresis separating tank and is interacted with enzyme connection label and light, directly observation or by position Camera directly over multichannel chromatoelectrophoresis separating plate or pick-up lens are observed and are recorded the surging image of protein, complete Protein chemistry electrochemiluminescent immunoassay is imaged；It is characterized in that：Described step 1 adopts protein chromatographic electricity as claimed in claim 1 Swimming and its in-situ chemical trace and immune imaging device, comprise the following steps that：1a）Close multi-channel switch, make described multichannel Path blockade between well and described track, is divided to described multichannel chromatoelectrophoresis by described chromatography liquid channel Chromatoelectrophoresis buffer solution required for adding in groove, adding in described protein well needs the protein of chromatographic analysis Sample, adds electrophoretic buffer, 1b in electrophoresis tank）Open multi-channel switch, make described multi-channel loading hole and described electrophoresis Path between swimming lane is opened, and adds suitable voltage simultaneously, make institute on left and right two described multichannel chromatoelectrophoresis handover modules State the protein example in protein well and move in the electric field in described multichannel chromatoelectrophoresis separating tank and separated.

7. protein chromatographic electrophoresis according to claim 6 and its in-situ chemical trace and immune imaging method, its feature It is：Described step 1 adopts protein chromatographic electrophoresis as claimed in claim 2 and its in-situ chemical trace and immune imaging dress Put, comprise the following steps that：1a）Close multi-channel switch, make the path between described multi-channel loading hole and described track Close, the chromatoelectrophoresis buffering required for being added in described multichannel chromatoelectrophoresis separating tank by described chromatography liquid channel Liquid, adding in described protein well needs the protein example of chromatographic analysis, adds electrophoretic buffer in electrophoresis tank, 1b）Open multi-channel switch, so that the path between described multi-channel loading hole and described track is opened, simultaneously left and right Add suitable voltage on two described multichannel chromatoelectrophoresis handover modules, so that the protein example in described protein well is existed The original position of described multichannel chromatoelectrophoresis separating tank, 1c is moved in electric field）Close multi-channel switch, make described multichannel Path blockade between well and described track；Meanwhile, by described chromatography liquid channel by required chromatographic flow phase Input multichannel chromatoelectrophoresis separating tank, makes protein example be separated in described chromatoelectrophoresis medium.

8. protein chromatographic electrophoresis according to claim 7 and its in-situ chemical trace and immune imaging method, its feature It is：Described step 4）Comprise the following steps that：4）Close multi-channel switch, make described multi-channel loading hole and described track Between path blockade, by described chromatography liquid channel sequentially input buffering in described multichannel chromatoelectrophoresis separating tank Liquid, the SA of the first antibody that one or more are capable of identify that some protein, fluorescence or enzyme connection mark.

9. multifunctional protein chromatoelectrophoresis and its recovery method it is characterised in that：Protein layer according to claim 1 Analysis electrophoresis and its in-situ chemical trace and immune imaging device, comprise the steps：1a）Close multi-channel switch, make described many Path blockade between passage well and described track, chromatographs electricity by described chromatography liquid channel to described multichannel Chromatoelectrophoresis buffer solution required for adding in swimming separating tank, adding in described protein well needs the egg of chromatographic analysis White matter sample, adds electrophoretic buffer, 1b in electrophoresis tank）Open multi-channel switch, make described multi-channel loading hole with described Path between track is opened, and adds suitable voltage on left and right two described multichannel chromatoelectrophoresis handover modules simultaneously, Make the protein example in described protein well move in the electric field in described multichannel chromatoelectrophoresis separating tank to obtain Separate；1c）Close multi-channel switch, make the path blockade between described multi-channel loading hole and described track；Meanwhile, lead to Cross described chromatography liquid channel and required chromatographic flow phase is inputted multichannel chromatoelectrophoresis separating tank, make the protein sample after separating Product are sequentially exported by the liquid-phase outlet of right multichannel chromatoelectrophoresis handover module and are collected respectively with collector.

10. multifunctional protein chromatoelectrophoresis method it is characterised in that：Protein chromatographic electrophoresis according to claim 2 And its in-situ chemical trace and immune imaging device, comprise the steps：1a）Adjust gas pressure in described pneumatic switch slot, So that described jettron film is under barotropic state, close logical between described multi-channel loading hole and described chromatography liquid channel Road, the chromatoelectrophoresis buffering required for being added in described multichannel chromatoelectrophoresis separating tank by described chromatography liquid channel Liquid, adding in described protein well needs the protein example of chromatographic analysis, adds electrophoretic buffer in electrophoresis tank, 1b）Adjust gas pressure in described pneumatic switch slot, so that described jettron film is under negative pressure state, open described multichannel Path between well and described chromatography liquid channel, simultaneously in left and right two described multichannel chromatoelectrophoresis handover modules Above add suitable voltage, make the protein example in described protein well move to described multichannel chromatoelectrophoresis in the electric field Separated in separating tank.