CN103439506A - Rapid detection device and method for interaction of biomacromolecules - Google Patents
Rapid detection device and method for interaction of biomacromolecules Download PDFInfo
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- CN103439506A CN103439506A CN2013103494687A CN201310349468A CN103439506A CN 103439506 A CN103439506 A CN 103439506A CN 2013103494687 A CN2013103494687 A CN 2013103494687A CN 201310349468 A CN201310349468 A CN 201310349468A CN 103439506 A CN103439506 A CN 103439506A
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Abstract
The invention relates to the field of bioengineering and aims to provide a rapid detection device and method for interaction of biomacromolecules. The device comprises a bottom plate, wherein the surface of the bottom plate is provided with n detection units, and n is an integral number greater than or equal to 1; each detection unit comprises a reaction hole, a detection hole and a micropipe for connecting the reaction hole and the detection hole; each detection hole is connected to a negative pressure device interface through the corresponding micropipe; the diameter of each micropipe is 0.1 to 2 mm; each reaction hole adopts an open structure and is used for dropping in a reactant; each detection hole adopts a closed structure and a magnetic metal wire is arranged in each detection hole; the direction of each metal wire is consistent with that of the micropipe in the corresponding detection unit. According to the invention, the interaction of antigen-antibodies or two biomacromolecules can be directly observed and the reaction condition can be judged by naked eyes or an instrument only in 1 to 2 minutes. The device is suitable for all biomolecules detected by adopting the ELISA (enzyme-linked immuno sorbent assay) principle and has the characteristics of high efficiency, simplicity, rapidness, convenience, wide application and the like.
Description
Technical field
The invention belongs to bioengineering field, be specifically related to a kind of device and method for detection of interaction of biomacromolecules.
Background technology
Enzyme linked immunosorbent assay (Enzyme-Linked Immunosorbent Assays, ELISA) from 1971 by Engvall and Perlman report since, in immunology and biochemical field, be widely used.Along with the continuous renewal of the updating of method, microplate reader, in the context of detection of antigen and antibody, ELISA is still current laboratory and detects one of technology the most frequently used in analyzing.The ultimate principle of ELISA method does not affect immunologic competence separately after being antigen and antibody molecule covalent bond.By antigen coated, seal, add primary antibodie, add ELIAS secondary antibody, the step such as substrate colour developing, judge the specificity of antigen and antibody response, determine the concentration of antibody or antigen by the depth of color reaction.This kind of chromogenic reaction can be carried out quantitative measurement by ELISA detector (microplate reader), makes the ELISA method become a kind of not only special but also responsive detection method.Yet the ELISA reaction needs a plurality of steps from antigen coated to the substrate colour developing, needs in time within 4-6 hour, even spend the night.Current business-like ELISA detection kit, the ELISA Plate that general employing contains pre-coated antigen or antibody, but on detection time, still need 2-4 hour.
Therefore, current detection technique need to solve fast detecting reaction problem from the time.For this reason, the immune colloidal gold chromatography technology starts to come out.The chromatography detection technique comes across 1988 the earliest, is a kind of pregnant inspection chromatograph test strip that Unipath company utilizes nano-colloid gold grain Development and Production, uses and can go out testing result in prefabricated test strips 3-5 minute.After this, immune colloid gold is widely used and develops in quick detection reagent.But the equipment such as the collaurum making of the different nano particles of colloidal gold technique needs, metal spraying, point sample, and the chromatography reaction is often had various reactions and is caused with ELISA.The antigen that can occur in ELISA and antibody response but do not occur in colloid gold chromatographic test paper strip.
Therefore, in order to solve long shortcoming of ELISA reaction time, can solve again the low sensitivity problem usually occurred in immunochromatography simultaneously.The invention provides a solution, that is: the device of a kind of fast detecting antigen and antibody response (or interaction of biomacromolecules).
Summary of the invention
The technical problem to be solved in the present invention is, overcomes deficiency of the prior art, and a kind of device for fast detecting and method of interaction of biomacromolecules is provided.
For the technical solution problem, solution of the present invention is:
A kind of device for fast detecting of interaction of biomacromolecules is provided, comprises base plate, be provided with n detecting unit on the surface of base plate, the integer that n is >=1; Each detecting unit comprises a reacting hole, detection a hole and the microtubule that is connected both; Each detects Kong Jun and is connected to the negative pressure device interface by microtubule; The diameter of described microtubule is 0.1～2 millimeter; Described reacting hole is opened type, for splashing into reactant; Described detection hole is closed type, and there is a magnetic metal wire of tool its inside, and metal wire is consistent with the direction of microtubule in this group detecting unit.
In the present invention, described base plate is the base plate of the inert materials such as plastics, pottery or tygon.
In the present invention, described microtubule is the pipeline of tygon, propylene or butylenes plastic material.
In the present invention, the capacity of described reacting hole is 10～500 microlitres.
In the present invention, described detecting unit has two at least, each detecting unit mutually side by side and be arranged in parallel, its reacting hole and detect the two ends of hole apportion base plate and arrange and embark on journey.
In the present invention, described negative pressure device interface is for connecting the interface of peristaltic pump or rubber pipette bulb.
The present invention also further provides a kind of method for quick of the interaction of biomacromolecules based on aforementioned means, comprises the following steps:
(1) according to the product operation instruction of immunomagnetic beads, the first biomacromolecule is coupled on immunomagnetic beads, then adds successively the second biomacromolecule of material to be detected and at least one mark; This process has two kinds of implementations: complete mixing in test tube after, reactant liquid is added to reacting hole, or add reactant liquid successively in reacting hole;
Described the first biomacromolecule is any one in protein, polysaccharide, polypeptide, nucleic acid or chemicals, and described material to be detected and the second biomacromolecule are any one in protein, polypeptide, polysaccharide, toxin, chemicals or nucleic acid.Wherein, the second biomacromolecule can be combined with material to be detected; The label that is used for the second biomacromolecule is any one of collaurum, fluorescent material, chemiluminescent material or upper forwarding luminescent material;
(2) start peristaltic pump, make the reactant liquid flow detection hole in reacting hole; If be coupled to the second biomacromolecule of the first biomacromolecule on magnetic bead, material to be detected, mark, interaction has occurred, reactant liquid is adsorbed on detection line at the detection Kong Shihui that flows through; The change color of magnet-wire can be detected by naked eyes, fluorescence detector, chemiluminescence detector or upper forwarding photodetector.
In the present invention, add in the reactant liquid in reacting hole, the concentration that is coupled to the first biomacromolecule on magnetic bead is 0.1～1 ug/ml; The concentration of the second biomacromolecule of mark is determined according to its manufacturer's recommendation.
The characteristic that the present invention utilizes the immunomagnetic beads of activation to react with biomacromolecule, by after the first biomacromolecule and immunomagnetic beads coupling, then seal unreacted avtive spot.Then the second biomacromolecule that adds material to be detected and mark.Wherein, thing energy while to be detected and the first biomacromolecule and the second biomacromolecule covalent bond, and formation is with the complex of " magnetic ", when reactant is flowed through the detection hole, can be adsorbed on the magnet wire detected in hole, and can be with the naked eye, fluorescence, chemiluminescence or on turn light-emitting appearance and detect.Use which kind of instrument to detect the label that depends on the second biomacromolecule.If label is collaurum, reactant compound naked eyes are visible.If label is fluorescent material, the reaction compound can use the fluorescence detector with optical filter to detect.As long as conditions being possessed, only need 1-2 minute from being reacted to detection.Because reaction is to carry out in liquid, therefore kept the immunologic competence of each reactive material in conventional ELISA, thereby realized the purpose of fast detecting.
Compared with prior art, the invention has the beneficial effects as follows:
The present invention can directly observe the interaction between antigen-antibody or two biomacromolecules, only needs 1-2 minute, can judge response situation by naked eyes or instrument.
This device is applicable to all biomolecule that the ELISA principle detects.
With respect to background technology, the characteristics such as that the present invention has is efficient, simple and direct, quick, convenient, applicable wide.
The accompanying drawing explanation
Fig. 1 is square (2x8 hole) of the present invention reaction unit schematic diagram.
Reference numeral in figure is: reacting hole 1; The metal wire 2 of magnetic; Detect hole 3; Peristaltic pump interface 4.Wherein, the reaction compound will be adsorbed on " metal wire of magnetic ".
Embodiment
Inventive principle is introduced:
Interaction of biomacromolecules device for fast detecting in the present invention, the principle of foundation is by the specific adsorption between immunomagnetic beads and magnet, and the covalent bond principle of antigen and antibody is made.By one of them biomacromolecule and immunomagnetic beads coupling.Another one biomacromolecule and enzyme, fluorophor, chemiluminescent groups or the coupling of upper forwarding light group.If two biomacromolecules directly or indirectly interact, can form the magnetic compound of tool.Thereby be adsorbed when flowing through magnet wire.
In the present embodiment, as shown in Figure 1, pick-up unit is square, and reacting hole is 2x8 hole (quantity is increased and decreased per sample in actual use), is arranged in parallel, and does not interfere with each other.Detect hole 3 corresponding one by one with reacting hole 1, detect between hole 3 and detection hole 1 and also do not interfere with each other.Be provided with a peristaltic pump interface 4 on the interconnective microtubule of detection hole 3 one side.Liquid in reacting hole 1 can be sucked and detects in hole 3 by the peristaltic pump negative pressure, and finally flow into waste fluid container.In the present invention, the device that also available rubber pipette bulb etc. can produce negative pressure replaces peristaltic pump, and its fundamental purpose is to promote liquid to flow through smoothly to detect hole.
Base plate is the inert materials such as plastics, pottery, tygon, and this material is not combined with biomacromolecules such as protein, polypeptide, polysaccharide, toxin, chemicals, nucleic acid, also with magnetisable material, " magnetic is combined " reaction does not occur.The capacity of reacting hole 1 is 10 microlitre to 500 microlitres, open.The microtubule material in coupled reaction hole 1 and detection hole 3 is tygon, propylene or butylenes plastic, and the microtubule diameter is 0.1 millimeter to 2 millimeters.Detecting hole 3 is closed type, only comprises the passage that liquid flows through, and capacity depends on the microtubule diameter.
Before detecting analysis, by the immunomagnetic beads coupling of first biomacromolecule and activation, conjugate can be placed on room temperature, 4 according to its character
oc or-20
oc saves backup.Then seal unreacted avtive spot on immunomagnetic beads with confining liquid, then the magnetic bead conjugate is mixed successively with the second biomacromolecule of material to be checked, mark, add in reacting hole 1.To install with peristaltic pump and be connected, start peristaltic pump, make liquid flow (or using rubber pipette bulb, generation suction).Liquid will be flowed through and detect hole 3 from reacting hole 1.By naked eyes, fluorescence detection device, chemiluminescence detecting or upper forwarding optical detection device, detected.Positive reaction, should present color in detection hole 3, and distribute along the metal wire of magnetic.
In the present invention, the product of the common commodity in use of the second biomacromolecule of mark, concrete reaction density determines according to manufacturer's recommendation concentration, different manufacturer's recommending datas slightly change.
Following embodiment can make the technician of this professional skill field more fully understand the present invention, but does not limit the present invention in any way.
Embodiment 1
1. the device for fast detecting of interaction of biomacromolecules is made
With reference to Fig. 1 custom mold, wherein die size: long x is wide=12x6 centimetre; Material is vinyon.The size of reacting hole 1: the wide x of long x is dark=0.7x0.5x0.3 centimetre, for open, can be used for application of sample.Detecting hole 3 measure-alike with reacting hole 1, but be closed, is only the metal wire 2 of magnetic for inlay one along the microtubule direction, size: the wide x of long x is high=and 0.5x0.1x0.1 centimetre.Reacting hole 1 is arranged in parallel with detecting hole 3, at a distance of 4 centimetres, by microchannel, is connected.0.1 centimetre of microchannel diameter.After all device for cleaning pipelines are crossed the detection hole, 1 centimetre of extension gathers to the peristaltic pump access port.Microchannel is Mold Making, one-shot forming.
2. application
The application of this device can be understood as a kind of immunomagnetic beads adsorption technology based on the ELISA principle.Current commercial immunomagnetic beads surface is containing amino, carboxyl, silica-based, Streptavidin, aldehyde radical or sulfydryl isoreactivity group.According to shop instruction by biomolecule and immunomagnetic beads coupling.Following program with the magnetic bead of Much's bacillus outer membrane protein OmpA and NHS activation (Pierce company, article No. 88826, can with contain amino biomolecule in conjunction with) coupling be example, set forth coupling process and the detection method of immunomagnetic beads and OmpA albumen:
Get 300 microlitre magnetic beads and be placed in 1.5 milliliters of centrifuge tubes, vertically be positioned on the base that contains magnet, draw supernatant and abandon.Add 1 mM hydrochloric acid of 1 milliliter of precooling, light shaking mixes, and vertically is positioned on the base that contains magnet, draws supernatant and abandons.(annotate: protein concentration is the 0.1-2 mg/ml, with the 50 mM borates of pH 8.5, dissolves to add immediately the OmpA albumen of 300 microlitre purifying; Perhaps with the damping fluid that does not contain amino pH 7-9, dissolve; Can not contain Tris or glycocoll in damping fluid), light shaking 30 seconds.Be placed on the rotation vortex mixer room temperature or 4
oc reaction 1 hour.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add 1 milliliter of 0.1M glycocoll (pH 2.0), concussion mixes 15 seconds.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.With 0.1M glycocoll (pH 2.0) cyclic washing magnetic bead 2 times.Add 1 milliliter of ultrapure water, concussion mixes 15 seconds.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add the reactive group of 1 milliliter of cancellation damping fluid (3M monoethanolamine, pH 9.0) sealing magnetic bead surfaces, after concussion mixes 30 seconds, room temperature reaction is 2 hours.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add 1 milliliter of ultrapure water, mix.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add 1 milliliter of storage liquid (50 mM borates, pH 8.5,0.05% Sodium azides), concussion mixes 15 seconds.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.With storage liquid washing 2 times.Finally with 300 microlitre storage liquid suspension magnetic bead protein conjugates, in 4
oc saves backup (now OmpA albumen-magnetic bead conjugate final concentration is 10 mg/ml).
By OmpA albumen-magnetic bead phosphate buffer (10mM for conjugate, pH 7.4) be diluted to 1 ug/ml, get 50 microlitres and be placed in 1.5 milliliters of centrifuge tubes, then add 50 microlitres mycobacterium tuberculosis antibody to be detected, then the Protein A/G(that adds 50 microlitre FITC marks also can be with marks such as HRP or collaurum or upper forwarding light particles).Light shaking mixes, room temperature reaction 5 minutes.100 microlitre response samples are dripped in the reacting hole 1 in the device for fast detecting of interaction of biomacromolecules.Connect peristaltic pump or rubber pipette bulb, make liquid from reacting hole 1 flow detection hole 3, end reaction liquid all flows out in waste liquid pool.
With the naked eye (be applicable to the Protein A/G of collaurum or HRP mark; If the HRP mark also needs to add o-phenylenediamine), fluorescence detection device (being applicable to the fluorescein-labeled Protein A/G such as FITC), upper forwarding optical detection device (being applicable to the Protein A/G of forwarding light particle marker) detect hole and response line whether occurs.
In response sample in splashing into reacting hole, the concentration of OmpA albumen-magnetic bead conjugate is 1 ug/ml, the detection lower limit line concentration of mycobacterium tuberculosis antibody to be detected is 0.001 ug/ml (all can detect higher than this concentration), the concentration of the Protein A/G of commercial FITC mark is 0.01 ug/ml (this concentration is recommended by the manufacturer, and the working concentration that different manufacturers recommend slightly changes).
Embodiment 2
1. the device for fast detecting of interaction of biomacromolecules is made, identical with embodiment 1.
2. application
The embodiment of the present embodiment is consistent with the principle of embodiment 1, following program be take nucleic acid as example, set forth magnetic bead (the Pierce company of itself and Streptavidin activation, article No. 88816, can with the biomolecule that contains biotin in conjunction with) coupling, set forth coupling process and the detection method of immunomagnetic beads and nucleic acid:
Get 50 microlitres (approximately 0.5 milligram) magnetic bead and be placed in 1.5 milliliters of centrifuge tubes, vertically be positioned on the base that contains magnet, draw supernatant and abandon.Add 1 milliliter of lavation buffer solution (10mM Tris-HCl, pH 7.0, containing 0.1% Tween-20) light shaking to mix, vertically be positioned on the base that contains magnet, draw supernatant and abandon.Add immediately the biotin labeled nucleic acid fragment of 300 microlitre (annotate: nucleic acid concentration is 2 ug/ml, dissolves with above-mentioned lavation buffer solution), light shaking 30 seconds.Be placed on the rotation vortex mixer room temperature or 4
oc reacts 1 hour or spends the night.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add 300 milliliters of lavation buffer solutions, draw supernatant and abandon, collect magnetic bead.Cyclic washing magnetic bead 2 times, finally use 300 milliliters of lavation buffer solution suspension magnetic beads.In 4
oc saves backup (now nucleic acid-magnetic bead conjugate final concentration is about the 1-2 ug/ml).
By nucleic acid-magnetic bead phosphate buffer (10mM for conjugate, pH 7.4) be diluted to 0.1 ug/ml, get 50 microlitres and be placed in 1.5 milliliters of centrifuge tubes, then add 50 microlitre the second biomolecule (little molecule toxin to be detected, concentration is 1 nanogram-1 ug/ml), add 50 microlitre toxin specific antibodies (monoclonal antibody, extension rate is 1:10000), two anti-(sheep anti-mouse antibodies of FITC mark, extension rate is 1:5000) that add again 50 microlitre marks.Light shaking mixes, room temperature reaction 5 minutes.100 microlitre response samples are dripped in the reacting hole 1 in the device for fast detecting of interaction of biomacromolecules.Connect peristaltic pump or rubber pipette bulb, make liquid from reacting hole 1 flow detection hole 3, end reaction liquid all flows out in waste liquid pool.Detecting step is with embodiment mono-.
In response sample in splashing into reacting hole, the concentration of nucleic acid-magnetic bead conjugate is 0.1 ug/ml, the detection least concentration of little molecule toxin to be detected is 1 nanograms/milliliter (all can detect higher than this concentration), the concentration of toxin specific antibody is 1 nanograms/milliliter, the concentration of the sheep anti-mouse antibody of commercial FITC mark is 0.01 ug/ml (this concentration is recommended by the manufacturer, and the working concentration that different manufacturers recommend slightly changes).
Embodiment 3
1. the device for fast detecting of interaction of biomacromolecules is made, identical with embodiment 1.
2. application
The embodiment of the present embodiment is consistent with embodiment 1 principle, following program be take antibody as example, sets forth magnetic bead (Pierce company, the article No. 88802 of itself and ProteinA/G activation, can with antibody molecule in conjunction with) coupling, set forth coupling process and the detection method of immunomagnetic beads and antibody:
Get 50 microlitres (approximately 0.5 milligram) magnetic bead and be placed in 1.5 milliliters of centrifuge tubes, vertically be positioned on the base that contains magnet, draw supernatant and abandon.Add 150 microlitre lavation buffer solutions (10mM Tris-HCl, pH 7.0, containing 0.1% Tween-20) light shaking to mix, vertically be positioned on the base that contains magnet, draw supernatant and abandon.Add 1 milliliter of lavation buffer solution, centrifuge tube vertically is positioned on the base that contains magnet, draw supernatant and abandon.Add immediately 500 microlitre samples (as: containing the serum of antibody, cell culture, ascites etc.), light shaking 30 seconds.Be placed on the rotation vortex mixer room temperature reaction 1 hour.Centrifuge tube vertically is positioned on the base that contains magnet, draws supernatant and abandon, collect magnetic bead.Add 300 milliliters of lavation buffer solutions, draw supernatant and abandon, collect magnetic bead.Cyclic washing magnetic bead 2 times, finally use 300 milliliters of lavation buffer solution suspension magnetic beads.In 4
oc saves backup.
Get 50 microlitre antibody-magnetic bead conjugate and be placed in 1.5 milliliters of centrifuge tubes, then add 50 microlitre marks, can with the little molecule toxin of antibody response on magnetic bead, add the aptamer of the FITC mark that can react with toxin, light shaking mixes again, room temperature reaction 5 minutes.100 microlitre response samples are dripped in the reacting hole 1 in the device for fast detecting of interaction of biomacromolecules.Connect peristaltic pump or rubber pipette bulb, make liquid from reacting hole 1 flow detection hole 3, end reaction liquid all flows out in waste liquid pool.Detecting step is with embodiment mono-.
In response sample in splashing into reacting hole, the concentration of antibody-magnetic bead conjugate is 0.5 ug/ml, the detection least concentration of toxin to be detected is 1 nanograms/milliliter (all can detect higher than this concentration), and the aptamer concentration of FITC mark is 0.1 ug/ml.
Be more than in conjunction with specific embodiments the son the present invention is done further describe.The technician of the industry should understand; the present invention is not restricted to the described embodiments; that in above-described embodiment and instructions, describes just illustrates principle of the present invention; under the premise without departing from the principles of the invention; the present invention and application thereof also have various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.
Claims (8)
1. the device for fast detecting of interaction of biomacromolecules, comprise base plate, it is characterized in that, is provided with n detecting unit on the surface of base plate, the integer that n is >=1; Each detecting unit comprises a reacting hole, detection a hole and the microtubule that is connected both; Each detects Kong Jun and is connected to the negative pressure device interface by microtubule; The diameter of described microtubule is 0.1～2 millimeter; Described reacting hole is opened type, for splashing into reactant; Described detection hole is closed type, and there is a magnetic metal wire of tool its inside, and metal wire is consistent with the direction of microtubule in this group detecting unit.
2. install according to claim 1, it is characterized in that, described base plate is the base plate of plastics, pottery or tygon inert material.
3. install according to claim 1, it is characterized in that, described microtubule is the pipeline of tygon, propylene or butylenes plastic material.
4. install according to claim 1, it is characterized in that, the capacity of described reacting hole is 10～500 microlitres.
5. according to the device described in claim 1 to 4 any one, it is characterized in that, described detecting unit has two at least, each detecting unit mutually side by side and be arranged in parallel, its reacting hole and detect the two ends of hole apportion base plate and arrange and embark on journey.
6. according to the device described in claim 1 to 4 any one, it is characterized in that, described negative pressure device interface is for connecting the interface of peristaltic pump or rubber pipette bulb.
7. the method for quick of the interaction of biomacromolecules based on the described device of claim 1, is characterized in that, comprises the following steps:
(1) according to the product operation instruction of immunomagnetic beads, the first biomacromolecule is coupled on immunomagnetic beads, then adds successively the second biomacromolecule of material to be detected and at least one mark; This process has two kinds of implementations: complete mixing in test tube after, reactant liquid is added to reacting hole, or add reactant liquid successively in reacting hole;
Described the first biomacromolecule is any one in protein, polysaccharide, polypeptide, nucleic acid or chemicals, described material to be detected and the second biomacromolecule are any one in protein, polypeptide, polysaccharide, toxin, chemicals or nucleic acid, and the second biomacromolecule of described mark can be combined with material to be detected; The mark that is used for the second biomacromolecule is any one of collaurum, fluorescent material, chemiluminescent material or upper forwarding luminescent material;
(2) start peristaltic pump, make the reactant liquid flow detection hole in reacting hole; If be coupled to the second biomacromolecule of the first biomacromolecule on magnetic bead, material to be detected, mark, interaction has occurred, reactant liquid is adsorbed on detection line at the detection Kong Shihui that flows through; The change color of magnet-wire can be detected by naked eyes, fluorescence detector, chemiluminescence detector or upper forwarding photodetector.
8. method according to claim 1, is characterized in that, adds in the reactant liquid in reacting hole, and the concentration that is coupled to the first biomacromolecule on magnetic bead is 0.1～1 ug/ml; The concentration of the second biomacromolecule of mark is determined according to its manufacturer's recommendation.
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| CN106706898A (en) * | 2016-11-23 | 2017-05-24 | 金莹 | Biological fluorescence detection system and detection method thereof |
| CN109613245A (en) * | 2019-01-09 | 2019-04-12 | 新疆医科大学 | A method of detection albumen-allinase conjugate targeting |
| CN109856392A (en) * | 2019-02-22 | 2019-06-07 | 武汉巴菲尔生物技术服务有限公司 | The device for fast detecting and method of interaction of biomacromolecules |
| CN111077301A (en) * | 2020-01-16 | 2020-04-28 | 浙江农林大学 | Micro-fluidic chip and method for detecting interaction between biomolecules |
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| CN103439506B (en) | 2015-04-15 |
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