CN103323603A - Protein covalent coupling method on surface of magnetic beads - Google Patents
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- CN103323603A CN103323603A CN2013102260704A CN201310226070A CN103323603A CN 103323603 A CN103323603 A CN 103323603A CN 2013102260704 A CN2013102260704 A CN 2013102260704A CN 201310226070 A CN201310226070 A CN 201310226070A CN 103323603 A CN103323603 A CN 103323603A
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Abstract
The invention discloses a protein covalent coupling method on the surface of magnetic beads. The method comprises the steps that: (1) the magnetic beads and an activating agent are subjected to co-incubation, such that activated magnetic beads are obtained; (2) the activated beads are subjected to co-incubation with protein, such that magnetic beads coupled with the protein are obtained; (3) the magnetic beads coupled with the protein are treated by using a blocking solution, such that surface active sites are blocked; and (4) the magnetic beads obtained in the step (3) are subjected to simultaneous vibration and sieving, and the sieved magnetic beads are collected. The method provided by the invention has the advantages of high protein coupling efficiency, good repeatability, and versatility. With the method, biological activities of protein coupled on the magnetic beads are not influenced. With the method provided by the invention, the obtained magnetic beads with covalently coupled protein can be used in fields such as immunoassay, biochemical detection, molecular detection, cell typing, and the like.
Description
Technical field
The present invention relates to a kind of at the magnetic bead surfaces method for covalent coupling protein.
Background technology
Magnetic bead claims again magnetic particle, is that a class diameter is at nanometer or micron-sized spherical compound substance.Magnetic bead is comprised of three parts, and core is small metal particles (iron oxide, tri-iron tetroxide), the outer evenly parcel of core one deck macromolecular material (such as polystyrene, Polyvinylchloride etc.), and outermost layer is function basic unit, has functional group, such as amino (NH
2), (COOH), hydroxyl (OH) etc. for carboxyl.
Magnetic bead is uniform-spherical, has superparamagnetism, makes magnetic bead move the purpose that reaches the separate targets thing to magnetic direction adding under the magnetic fields, and outermost chemical functional group is used to and the bioactive molecule couplings such as protein or nucleic acid.Magnetic bead after the coupling is widely used in the fields such as cell separation and purifying, cell typing, immune detection, separate nucleic acid, targeting drug release system.
The carboxyl magnetic bead, namely outermost layer has the carboxyl function group (magnetic bead COOH) is a class magnetic bead of comparatively commonly using.The coupling method of carboxyl magnetic bead and protein has a variety of at present.Because the difference on selection, activation condition, coupling condition and the coupling process of activator, cause that the magnetic bead of coupling protein matter exists that coupling efficiency is low, protein utilization rate is low, the autoagglutination of coupling magnetic bead, the non-specific absorption of magnetic bead are serious, can not be general etc. deficiency.
Summary of the invention
The purpose of this invention is to provide a kind of at the magnetic bead surfaces method for covalent coupling protein.
Provided by the invention at the magnetic bead surfaces method for covalent coupling protein, comprise the steps:
(1) magnetic bead and activator are hatched altogether the magnetic bead after obtaining activating;
(2) magnetic bead after the described activation and protein are hatched altogether, obtain the magnetic bead that coupling has described protein;
(3) there is the magnetic bead of described protein to process with confining liquid described coupling, the avtive spot of confining surface;
(4) sieving (to remove the magnetic particle of reunion) in the limit that vibrates, the magnetic bead limit that step (3) is obtained, collects the magnetic bead after sieving.
In the described step (1), described magnetic bead can be the carboxyl magnetic bead.In the described step (1), the magnetic bead surfaces after the described activation has the chemical group that contains N-maloyl imine linkage.In the described step (1), described activator can be EDC and NHS, or described activator can be EDC and Sulfo-NHS.In the described step (1), described reaction system of hatching altogether can be the MES damping fluid of pH4.0-7.0,0.01-0.1M.In the described step (1), when described activator was EDC and NHS, the initial concentration of described EDC can be 0.1-30mM, and the initial concentration of described NHS can be 0.3-100mM.In the described step (1), when described activator was EDC and Sulfo-NHS, the initial concentration of described EDC can be 0.1-30mM, and the initial concentration of described Sulfo-NHS can be 0.3-100mM.In the described step (1), the described time of hatching altogether can be 6-300 minute, is preferably 10-240 minute.In the described step (1), described temperature of hatching altogether specifically can be room temperature.
In the described step (2), described reaction system of hatching altogether can be the MES damping fluid of pH4.0-7.0,0.01-0.1M; In the described step (2), described condition of hatching altogether can be: 2-26 ℃, 2-20 hour.
In the described step (3), described confining liquid can be the damping fluid that contains the 0.2-5g/100mL closed protein, is preferably the damping fluid that contains 1-5g/100mL BSA.Described damping fluid can be MES damping fluid, PBS damping fluid or Tris damping fluid.The MES damping fluid is the 2-(morpholino) the ethyl sulfonic acid damping fluid.The PBS damping fluid is phosphate buffer.The Tris damping fluid is TRIS buffer.In the described step (3), the processing time of described " having the magnetic bead of described protein to process with confining liquid described coupling " can be 1-9 hour, is preferably 90-180 minute.In the described step (3), described temperature of hatching altogether specifically can be room temperature.
In the described step (4), described sieving can be 800 orders-1500 mesh sieve.Described sieving specifically can be stainless steel mesh.Described stainless steel mesh specifically can be 304 steel stainless steel mesh or 316 steel stainless steel mesh.In the described step (4), described sieve by liquid-phase system specifically can be the MES damping fluid of pH4.0-7.0,0.01-0.1M.
Described method also can comprise with step (4) obtain sieve after magnetic bead with the step of preserving 4 ℃ of preservations behind the liquid suspendible.Described preservation liquid is for containing 0.1-5g/100mL BSA and 0.1g/100mL NaN
3PH7-10,0.01-0.1MTris damping fluid.
In the described step (2), described protein specifically can be antibody, such as goat-anti fluorescein isothiocynate IgG antibody or thyroxine monoclonal antibody.
The magnetic bead product that the present invention also protects above arbitrary described method to prepare.
The present invention also protects the application of described magnetic bead product in the material of evaluation and/or separation and described protein bound.When described protein was antibody, described " with the material of described protein bound " was and the antigen of described protein specific bond or two anti-.
The present invention has following advantage:
(1) albumen coupling efficient is high; Use dual activator EDC and NHS activation magnetic bead, can make the activated carboxyl on the magnetic bead be in semisteady-state, not facile hydrolysis, after adding protein, form stable amido link with the amino of protein side chain, increased the utilization factor of magnetic bead and antibody, increased coupling efficiency;
(2) good reproducibility; By the step that vibration is sieved, removed the magnetic bead particles that produces aggegation, make the magnetic bead particles homogeneous, easily mixing is without reunion, and immune response efficient is improved, and avoids non-specific adsorption, has guaranteed the repeatability that immune response is good;
(3) has versatility; Step by sealing obtains the unreacted site of magnetic bead itself saturated, has greatly reduced the generation of non-specific absorption, has increased the versatility of magnetic bead;
(4) do not affect the biologic activity that is coupled at the protein on the magnetic bead; On the one hand, the mild conditions such as the temperature that activation and coupling reaction occur, ionic strength, pH can not cause protein denaturation; On the other hand, the steric configuration of protein is unaffected after protein and the magnetic bead coupling, still can keep its original biologically active.
The magnetic bead of the covalent coupling protein that method provided by the invention obtains can be used for the fields such as immune detection, biochemistry detection, Molecular Detection, cell typing.
Description of drawings
Fig. 1 EDC and NHS activated carboxyl magnetic bead reaction schematic diagram.
Fig. 2 activates rear magnetic bead and proteins react schematic diagram.
Fig. 3 magnetic bead front design sketch that sieves.
Fig. 4 magnetic bead rear design sketch that sieves.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is conventional method.Used test material among the following embodiment if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples all arranges repeated experiments three times, results averaged.Among the embodiment, by the protein concentration in the ultraviolet spectrophotometry detection solution.IgG antibody and monoclonal antibody are protein, so among the embodiment, the amount of IgG antibody and the amount of monoclonal antibody are all in protein content.
EDC, Chinese full name are 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides, and English full name is 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride.Sulfo-NHS, Chinese full name are the N-hydroxy thiosuccinimide, and English full name is N-hydroxysulfosuccinimide.NHS, Chinese full name are N-hydroxy-succinamide, and English full name is N-hydroxysuccinimide.
EDC: available from Sigma-Aldrich, CAS:25952-53-8CAT:E7750.Sulfo-NHS: available from Sigma-Aldrich, CAS:106627-54-7CAT:56485.NHS: available from Sigma-Aldrich, CAS:6066-82-6CAT:56480.Carboxyl magnetic bead colloidal solution: available from Merck, article No. is EM1-100/40.Goat-anti fluorescein isothiocynate antibody serum: available from PLA General Hospital the first animal used as test section of affiliated hospital, article No.: Y11005.The thyroxine monoclonal antibody: available from Meridia, CAT:MAT02-211.
Embodiment 1, carboxyl magnetic bead coupling goat-anti fluorescein isothiocynate antibody
One, the activation of carboxyl magnetic bead
The purpose of this step is: the chemical group that generates the inferior diamino of carbon containing in magnetic bead surfaces.
1, magnetic bead pre-service
Get the sedimentation 20 minutes under magnetic fields of 1000mg carboxyl magnetic bead colloidal solution, abandon supernatant, then with magnetic bead with the MES damping fluid washing of 50ml pH7.0,0.01M 1 time.
2, add EDC and NHS behind the abundant suspendible of MES damping fluid of the magnetic bead that step 1 is obtained 20ml pH7.0,0.01M, the concentration that makes EDC is 0.1mM, the concentration of NHS is 100mM, and room temperature oscillating reactions (30 rev/mins) 240 minutes obtains activating magnetic bead solution.
Two, carboxyl magnetic bead coupling goat-anti fluorescein isothiocynate antibody
1, gets 10ml goat-anti fluorescein isothiocynate antibody serum, obtain goat-anti fluorescein isothiocynate IgG antibody (goat-anti fluorescein isothiocynate IgG antibody total amount is greater than 200mg) with protein-G affinity column purifying.
2, the goat-anti fluorescein isothiocynate IgG antibody that step 1 is obtained is splined on Sephadex G25 chromatographic column and carries out desalination, adopts the MES buffer solution elution of pH7.0,0.01M, collects antibody-solutions (the antibody total amount is greater than 150mg).
3, coupling
The antibody-solutions that step 2 is obtained joins in the activation magnetic bead solution that step 1 obtains, jog mixing, 2 ℃ of oscillating reactionss (20 rev/mins) 20 hours.In this course of reaction, the generation covalent reaction of the N-maloyl imine linkage of activation and the amino of antibody, antibody by covalent coupling in magnetic bead surfaces.
4, sealing
Take into the liquid-phase system of step 3, sedimentation is 20 minutes under magnetic fields, abandons supernatant, adds the 100ml confining liquid, jog mixing, room temperature oscillating reactions (30 rev/mins) 90 minutes.
Confining liquid: the MES damping fluid that contains pH7.0, the 0.01M of 1g/100mL BSA.
5, sieve
Take into the liquid-phase system of step 4, sedimentation was abandoned supernatant after 20 minutes under magnetic fields; The MES damping fluid of magnetic bead with pH7.0,0.01M washed 3 times each 100ml; Magnetic bead being suspended in crossing 1500 order materials after the MES damping fluid of 100ml pH7.0,0.01M is the stainless steel sift of 304 steel, in the process of sieving, constantly vibration and repeatedly wash with the MES damping fluid of pH7.0,0.01M, until the agglomerated particle that residue can not be sieved on the compass screen surface, magnetic bead after sieving is answered the particle homogeneous behind the suspendible again, without aggegation.
6, dilution and preservation
Collect the magnetic bead solution after the sieving that step 5 obtains, sedimentation was abandoned supernatant after 20 minutes under magnetic fields; Magnetic bead is cleaned 3 times each 100ml with preserving liquid; Then magnetic bead is diluted to 5mg/ml with preservation liquid, the room temperature suspendible is 4 ℃ of preservations after 2 hours.
Preserve liquid: the pH7.0, the 0.01M Tris damping fluid that contain 0.1g/100mL BSA and 0.1g/100mL NaN3.
Three, the non-specific adsorption behind the carboxyl magnetic bead coupling goat-anti fluorescein isothiocynate antibody
For detection of non-specific adsorption is carcinomebryonic antigen (CEA) detection kit (Magnetism particulate immuno chemistry luminescence method) of Boao Biological Co., Ltd; production code member 330410, key component have antibody reagent (the CEA antibody of marked by fluorescein isothiocyanate and the CEA antibody of alkali phosphatase enzyme mark), CEA calibration object (comprising 0 concentration point and 5 gradient concentration points), CEA quality-control product (Q1, Q2), magnetic particle reagent, luminous substrate (enzymatic is luminous), cleaning concentrate.The kit principle of work: the CEA antibody of the CEA antibody of marked by fluorescein isothiocyanate, CEA standard items and alkali phosphatase enzyme mark forms the compound of " sandwich " structure, this compound is connected with magnetic particle by the effect of fluorescein isothiocynate with goat-anti fluorescein isothiocynate antibody, add luminous substrate behind the washing magnetic particle, enzymatic is luminous, can detect fluorescence.
1, replaces magnetic particle reagent in the kits with the magnetic beads of 5 preparations of step 2, operate to specifications, obtain the typical curve take CEA concentration and fluorescence intensity as coordinate.
2, the magnetic beads of 5 preparations of usefulness step 2 replace the magnetic particle reagent in the kit, the CEA antibody that does not add marked by fluorescein isothiocyanate, other operates to specifications, adopt the CEA standard items of 0 concentration, the fluorescence intensity reference standard curve that detects, the CEA concentration that obtains is the result of non-specific adsorption.Non-specific adsorption＜the 0.001ng/ml of the magnetic bead of 5 preparations of step 2.
Replace magnetic particle reagent in the kit with the magnetic beads of 5 preparations of step 2, adopt CEA quality-control product Q1 as detecting sample, by specification operates, the fluorescence intensity reference standard curve that detects, the CEA concentration that obtains is the fitting result of quality-control product Q1, and the coefficient of variation of 5 repeated experiments is 1.31%.
3, replace magnetic particle reagent in the kits with the magnetic beads of 4 preparations of step 2, operate to specifications, obtain the typical curve take CEA concentration and fluorescence intensity as coordinate.
4, the magnetic beads of 4 preparations of usefulness step 2 replace the magnetic particle reagent in the kit, the CEA antibody that does not add marked by fluorescein isothiocyanate, other operates to specifications, adopt the CEA standard items of 0 concentration, the fluorescence intensity reference standard curve that detects, the CEA concentration that obtains is the result of non-specific adsorption.The non-specific adsorption of the magnetic bead of 4 preparations of step 2 is 2.03ng/ml.
Replace magnetic particle reagent in the kit with the magnetic beads of 4 preparations of step 2, adopt CEA quality-control product Q1 as detecting sample, by specification operates, the fluorescence intensity reference standard curve that detects, the CEA concentration that obtains is the fitting result of quality-control product Q1, and the coefficient of variation of 5 repeated experiments is 8.63%.
Four, the activity behind the carboxyl magnetic bead coupling goat-anti fluorescein isothiocynate antibody
1, gets the magnetic bead of 5 preparations of step 2, after preserving the liquid gradient dilution, obtain each dilution suspension.
2, the dilution suspension that 30 μ l steps 1 is obtained and the anti-sheep IgG of 30 μ l alkali phosphatase enzyme mark rabbits antibody (grind grass Bioisystech Co., Ltd available from Shanghai, production code member: E030230) mixing, 37 ℃ of incubations 5 minutes.
3, sedimentation was abandoned supernatant after 2 minutes under magnetic fields, magnetic bead was washed 3 times each 300 μ l with cleaning fluid (available from Boao Biological Co., Ltd, one of kit component of production code member 330410).
4, lucifuge adds 200 μ l alkaline phosphatase catalytic luminescence substrates (Lumigen APS-5), detects luminous value.
When adopting the dilution suspension of higher concentration, the magnetic bead luminous value is consistent substantially (because the anti-sheep IgG of the relative alkali phosphatase enzyme mark rabbit of magnetic bead antibody is excessive state, the anti-sheep IgG of all alkali phosphatase enzyme mark rabbits antibody all can be adsorbed by magnetic bead), but obvious decline (this moment, magnetic bead was not enough to adsorb all anti-sheep IgG of alkali phosphatase enzyme mark rabbit antibody) can appear in the magnetic bead luminous value when adopting the dilution suspension of a certain low concentration, this the magnetic bead concentration reflection magnetic beads activity that luminous value obviously descends occurs, the lower magnetic beads activity of this concentration is better, the higher leveled concentration of concentration that obviously descends than this luminous value is optimum activity concentration and (for example dilutes that magnetic bead concentration is followed successively by 5 in the suspension, 4,3,2,1mg/ml, if luminous value begins obvious decline when the concentration of 2mg/ml, optimum activity concentration is 3mg/ml).The optimum activity concentration of magnetic beads with 5 preparations of step 2 is 1.5mg/ml.
Get the magnetic bead of 4 preparations of step 2, detect optimum activity concentration according to above identical step.The optimum activity concentration of the magnetic bead of 4 preparations of step 2 is 3.5mg/ml.
Embodiment 2, carboxyl magnetic bead coupling monoclonal antibody
One, the activation of carboxyl magnetic bead
1, magnetic bead pre-service
Get the sedimentation 10 minutes under magnetic fields of 200mg carboxyl magnetic bead colloidal solution, abandon supernatant, with the magnetic bead of sedimentation with the MES buffer solution for cleaning of pH4.0,0.1M 3 times, each 20ml.
2, add EDC and NHS behind the abundant suspendible of MES damping fluid of the magnetic bead that step 1 is obtained 20ml pH4.0,0.1M, the concentration that makes EDC is 30mM, and the concentration of NHS is 0.3mM, and room temperature oscillating reactions (20 rev/mins) 10 minutes is activation magnetic bead solution.
Two, carboxyl magnetic bead coupling monoclonal antibody
1, gets the thyroxine monoclonal antibody, be splined on Sephadex G25 chromatographic column, adopt the MES buffer solution elution of pH4.0,0.1M, collect the 4ml antibody-solutions.
2, coupling
The antibody-solutions that step 1 is obtained joins in the activation magnetic bead solution that step 1 obtains, jog mixing, room temperature oscillating reactions (20 rev/mins) 2 hours.
3, sealing
Take into the liquid-phase system of step 2, sedimentation is 10 minutes under magnetic fields, abandons supernatant, adds the 20ml confining liquid, jog mixing, room temperature oscillating reactions (20 rev/mins) 180 minutes.
Confining liquid: the MES damping fluid that contains pH4.0, the 0.1M of 5g/100mL BSA.
4, sieve
Take into the liquid-phase system of step 3, sedimentation was abandoned supernatant after 10 minutes under magnetic fields; With magnetic bead with 4.0, the MES damping fluid washing of 0.1M 3 times, each 20ml; Magnetic bead being suspended in crossing 800 order materials after the MES damping fluid of 20ml pH4.0,0.1M is the stainless steel sift of 316 steel, in the process of sieving, constantly vibrates and repeatedly washes with the MES damping fluid of pH4.0,0.1M, until remain the bulky grain that can not sieve on the compass screen surface.Magnetic bead after sieving is answered the particle homogeneous behind the suspendible again, without aggegation.
5, dilution and preservation
Magnetic bead solution after collection is sieved, sedimentation was abandoned supernatant after 20 minutes under magnetic fields; Magnetic bead is cleaned 3 times each 100ml with preserving liquid; Then magnetic bead is diluted to 5mg/ml with preservation liquid, the room temperature suspendible is 4 ℃ of preservations after 2 hours.
Preserve liquid: contain 5g/100mL BSA and 0.1g/100mL NaN
3PH10.0,0.1M Tris damping fluid.Three, the activity after the carboxyl magnetic bead coupling thyroxine monoclonal antibody
1, gets the magnetic bead of 4 preparations of step 2, after preserving the liquid gradient dilution, obtain each dilution suspension.
The dilution suspension that 2,30 μ l steps 1 is obtained and the thyroxine of 60 μ l alkali phosphatase enzyme marks (available from Boao Biological Co., Ltd, production code member: one of kit component of 330150) mixing, 37 ℃ of incubations 6 minutes.
3, sedimentation was abandoned supernatant after 2 minutes under magnetic fields, magnetic bead was washed 3 times each 300 μ l with cleaning fluid (available from Boao Biological Co., Ltd, one of kit component of production code member 330150).
4, lucifuge adds 200 μ l alkaline phosphatase catalytic luminescence substrates (Lumigen APS-5), detects luminous value.When adopting the dilution suspension of higher concentration, the magnetic bead luminous value is consistent substantially (because the thyroxine of the relative alkali phosphatase enzyme mark of magnetic bead is excessive state, the thyroxine of all alkali phosphatase enzyme marks all can be adsorbed by magnetic bead), but obvious decline (this moment, magnetic bead was not enough to adsorb the thyroxine of all alkali phosphatase enzyme marks) can appear in the magnetic bead luminous value when adopting the dilution suspension of a certain low concentration, this the magnetic bead concentration reflection magnetic beads activity that luminous value obviously descends occurs, the lower magnetic beads activity of this concentration is better, the higher leveled concentration of concentration that obviously descends than this luminous value is optimum activity concentration and (for example dilutes that magnetic bead concentration is followed successively by 5 in the suspension, 4,3,2,1mg/ml, if luminous value begins obvious decline when the concentration of 2mg/ml, optimum activity concentration is 3mg/ml).The optimum activity concentration of magnetic beads with 4 preparations of step 2 is 0.5mg/ml.
Get the magnetic bead of 3 preparations of step 2, detect optimum activity concentration according to above identical step.The optimum activity concentration of the magnetic bead of 3 preparations of step 2 is 2mg/ml.
Adopt Sulfo-NHS to replace NHS to carry out respectively the concrete experiment of embodiment 1 and embodiment 2, obtained identical result.
Claims (10)
1. one kind at the magnetic bead surfaces method for covalent coupling protein, comprises the steps:
(1) magnetic bead and activator are hatched altogether the magnetic bead after obtaining activating;
(2) magnetic bead after the described activation and protein are hatched altogether, obtain the magnetic bead that coupling has described protein;
(3) there is the magnetic bead of described protein to process with confining liquid described coupling, the avtive spot of confining surface;
(4) sieving in the limit that vibrates, the magnetic bead limit that step (3) is obtained, collects the magnetic bead after sieving.
2. the method for claim 1, it is characterized in that: in the described step (1), described magnetic bead is the carboxyl magnetic bead.
3. method as claimed in claim 2, it is characterized in that: in the described step (1), the magnetic bead surfaces after the described activation has the chemical group that contains N-maloyl imine linkage.
4. method as claimed in claim 2 or claim 3, it is characterized in that: in the described step (1), described activator is EDC and NHS, or described activator is EDC and Sulfo-NHS.
5. method as claimed in claim 4 is characterized in that:
In the described step (1), the MES damping fluid that described reaction system of hatching altogether is pH4.0-7.0,0.01-0.1M;
In the described step (1), when described activator was EDC and NHS, the initial concentration of described EDC was 0.1-30mM, and the initial concentration of described NHS is 0.3-100mM;
In the described step (1), when described activator was EDC and Sulfo-NHS, the initial concentration of described EDC was 0.1-30mM, and the initial concentration of described Sulfo-NHS is 0.3-100mM;
In the described step (1), the described time of hatching altogether is 6-300 minute, is preferably 10-240 minute.
6. such as arbitrary described method in the claim 1 to 5, it is characterized in that: in the described step (2), the MES damping fluid that described reaction system of hatching altogether is pH4.0-7.0,0.01-0.1M; In the described step (2), described condition of hatching altogether is: 2-26 ℃, 2-20 hour.
7. such as arbitrary described method in the claim 1 to 6, it is characterized in that: in the described step (3), described confining liquid is the damping fluid that contains the 0.2-5g/100mL closed protein; Described damping fluid is MES damping fluid, PBS damping fluid or Tris damping fluid; In the described step (3), the processing time of described " having the magnetic bead of described protein to process with confining liquid described coupling " is 1-9 hour.
8. such as arbitrary described method in the claim 1 to 7, it is characterized in that: in the described step (4), described sieving as crossing 800 orders-1500 mesh sieve.
9. the magnetic bead product that arbitrary described method prepares in the claim 1 to 8.
10. the application of the described magnetic bead product of claim 9 in the material of evaluation and/or separation and described protein bound.
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| CN110058022A (en) * | 2019-05-29 | 2019-07-26 | 山东博科生物产业有限公司 | Carcinoembryonic Antigen CEA immue quantitative detection reagent box and its application |
| WO2021109684A1 (en) * | 2019-12-06 | 2021-06-10 | 南京师范大学 | Capturing magnetic bead for capturing weakly interacting protein molecule by employing photoaffinity covalent attachment, and preparation method therefor and use thereof |
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| CN113390837B (en) * | 2021-05-26 | 2023-09-29 | 安徽省昂普拓迈生物科技有限责任公司 | Method for detecting coupling efficiency of magnetic bead protein |
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