CN106769336B - Utilize the method for the micro- cage capture protein of hydrogel - Google Patents
Utilize the method for the micro- cage capture protein of hydrogel Download PDFInfo
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Abstract
The present invention relates to a kind of methods using the micro- cage capture albumen of hydrogel, it includes that (1) makes hyperbranched poly glycerin solution, double mercapto-polyglycol aqueous solutions, is mixed to get mixed liquor on micro-fluidic chip with markd protein peptides monomer solution to be captured, mix mixed liquor, oil phase substance on micro-fluidic chip, obtain microlayer model, in microlayer model, hyperbranched poly glycerine and double mercapto-polyglycols occur sulfydryl-alkene click chemistry and react to form hydrogel, obtain microgel；(2) carrying out culture to microgel makes albumen peptide monomer therein occur to fold aggregation；(3) tretolite is added to destroy the oily phase outer layer of microgel and be cleaned.The present invention can only capture the high protein oligomer of false folding degree and fiber, to carry out for Journal of Sex Research.Meanwhile the formation of hydrogel in the process will not have an impact protein structure, keep testing result more acurrate without heating or ultraviolet irradiation.
Description
Technical field
Related protein such as beta-amyloid protein, tau egg are generated with neurodegenerative disease the present invention relates to a kind of
The catching method of white, a- synapse nucleoprotein etc..The catching method can for the aggregation situation of accurate analysis and research protein to
Necessary condition is provided further to develop the therapeutic agent of these diseases.
Background technique
For world population in continuous aging, according to the statistics of the World Health Organization, there are 35,600,000 dementia patients in the current whole world, arrives
This number will double the year two thousand thirty, and the year two thousand fifty, this number will be increased to three times or more.Dementia type includes alzheimer '
The neurodegenerative diseases such as silent disease, Parkinson's disease, Huntington's chorea.Generally believe the protein of a large amount of false foldings, such as
It is mind that beta-amyloid protein (amyloid beta), Protein tau, a- synapse nucleoprotein (α-synuclein) are assembled in the cell
The material base generated through degenerative disease.Therefore during exploitation is directed to the therapeutic agent of these diseases, one is needed
Kind method easy to use captures these albumen, and monitors influence of these substances for protein aggregation in real time.
Hydrogel is the three-dimensional polymer space net structure formed by the polymer chain being crosslinked.It is not soluble in water but hydrophilic
Property can accommodate a large amount of water.Furthermore, it is possible to change aperture by the molecular weight for adjusting polymer concentration and precursor.Its
Drug molecule can be wrapped in polymer network by inner porosity.In addition, this structure can also be used as cell encapsulation
Bracket: its nanoscale aperture can permit it is subsequent supply nutrients to cell, but prevent larger substance such as antibody or enzyme simultaneously
Infiltration.In addition, the biocompatibility of hydrogel can preferably simulated albumin matter fold natural environment.
Hydrogel is generally divided into physical gel and two kinds of chemical gel.Wherein physical gel includes heat-convertible gel,
It is in stable gel state under room temperature, but solution can be changed into after heating.But wherein may to influence protein anti-for the change of temperature
Speed is answered, protein structure is destroyed.And chemical gel using it is more include containing methacrylate or acrylate group
Hyperbranched poly glycerine (hPG) and polyethylene glycol (PEG).By the way that these macromonomers are exposed to together with photoinitiator
Quick cross-linking is carried out under ultraviolet light, or is caused free radical using heating means and polymerize.To sum up, heating or purple are either utilized
The method of outside line irradiation, will affect albumen qualitative response, so that testing result is inaccurate.In addition, existing conventional physics
The not only wrong folding degree that gel and chemical gel catching method capture high protein oligomer and fiber, can also capture
The low small protein of false folding degree and part small size monomer, protein oligomerization that can not be high to false folding degree
Object and fiber are carried out for Journal of Sex Research.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and provide a kind of new protein capture side
Method, this method protein oligomer high to false folding degree and fiber can carry out specific aim capture, meanwhile, this method is not necessarily to
Heating or ultraviolet irradiation are relied on so as to avoid impacting albumen qualitative response, it is hereby achieved that more accurately analysis
As a result.
In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:
A method of albumen is captured using the micro- cage of hydrogel comprising following steps:
(1) make hyperbranched poly glycerin solution, double mercapto-polyglycol aqueous solutions, with markd protein peptides to be captured
Monomer solution is mixed to get mixed liquor on micro-fluidic chip, then makes the mixed liquor, oil phase substance in the micro-fluidic core
Mixing on chip obtains microlayer model, and in the microlayer model, sulfydryls-occur for the hyperbranched poly glycerine and double mercapto-polyglycols
The click chemistry of alkene reacts to form hydrogel, obtain outer layer be oily phase internal layer be the microgel containing albumen peptide solution；
(2) carrying out culture to the microgel makes albumen peptide monomer therein occur to fold aggregation, obtains outer layer as oily phase
Internal layer contains the microgel of the polymeric protein of different folding degree；
(3) tretolite is added in by the system of step (2) to destroy the oily phase outer layer and be cleaned to remove
It goes in the microgel less than the polymeric protein in the microgel duct or unpolymerized albumen to get the hydrogel for capturing albumen
Micro- cage, the micro- cage of hydrogel for capturing albumen can be used for subsequent detection and analysis.
Further, in step (1), the micro-fluidic chip includes chip body, is formed on chip body and one
End, which intersects, constitutes the first microchannel, the second microchannel and the third microchannel of the first focal zone, is formed on chip body
And portion intersects and constitutes the 4th microchannel, the 5th microchannel and the 6th microchannel of the second focal zone at one end, and is formed in
The 7th microchannel on chip body, wherein the other end of the 4th microchannel is connected to first focal zone, described
One end of seven microchannels is connected to second focal zone, and hyperbranched poly glycerin solution, double mercapto-polyglycols is water-soluble
Liquid is injected separately into first microchannel, the second microchannel and third microchannel with markd albumen peptide solution to be captured, and three
Person is mixed to get the mixed liquor in the first focal zone, injects the oil phase thing into the 5th microchannel and the 6th microchannel respectively
Matter, mixed liquor flow to the second focal zone through the 4th microchannel, infuse in two focal zones with out of the 5th microchannel and the 6th microchannel
The oil phase substance entered is mixed to form the microlayer model.
Further, filter (filtering accuracy is for example, about 1 micron) is arranged in the liquid feeding end in each microchannel, to prevent
Only particulate matter that may be present causes channel blockage.
Further, the method also includes making micro-fluidic chip with Soft lithograph legal system, which includes as follows
Several steps:
I) by designed first microchannel, the second microchannel, third microchannel, the 4th microchannel, the 5th microchannel with
And the 6th microchannel be printed on Acetate Film, and Acetate Film is placed at the top of the silicon wafer coated with photoresist, process is ultraviolet
Light irradiation, obtains chip template；
Ii) polydimethylsiloxane prepolymer object and its crosslinking agent are poured in the chip template, is solidified, will be consolidated
The dimethyl silicone polymer mold of change removes chip template, and carries out pressure piercing to the entrance of each microchannel, later
Each microchannel is sealed with the substrate of surfacing；
Iii each micro channel) is injected into surface hydrophobicity agent and is stood, so that microchannel surface is hydrophobic up to described micro-fluidic
Chip.
Further, in step (1), the markd protein peptides monomer solution to be captured of band can be by first using organic
Fluorescein carries out fluorescent marker to the end N- of albumen peptide monomer side chain, is then dissolved in 0.5%~2% ammonium hydroxide and prepares in advance
Standby protein peptides monomer solution is finally diluted to setting concentration with the buffer of pH 7-8 and obtains.
According to the present invention, above-mentioned organic fluorescence pigment includes but is not limited to Alexa Fluor405, Alexa-350, AMCA-
X、Alexa-488、FITC、HiLyte Flour488、Alexa-430、Alexa-555、HiLyte Plus555、
DyLight549、HiLyte Fluor555、Alexa Fluor 546、Alexa-546、Alexa-568、AlexaFlour
594, HiLyte Fluor TR (622), Alexa-633 etc..
Further, in step (1), double mercapto-polyglycol aqueous solutions by by 2- iminothiolane hydrochloride with
Polyethylene glycol diamine reacts in the buffer solution that pH is 7-8 to be obtained.
Further, in step (1), the albumen peptide monomer is people's amyloid beta 1-40, people's amyloid beta 1-
42, Protein tau or a- synapse nucleoprotein.
Further, in step (1), the oil phase substance is preferably selected from fluorination liquid, mineral substance oil, vegetable oil and fat
One of acid or a variety of combinations.
Preferably, in step (1), the flow velocity of the hyperbranched poly glycerine and double mercapto-polyglycol aqueous solutions is respectively
The flow velocity of the markd protein peptides monomer solution to be captured of micro- l/h of 10-400, the band is micro- l/h of 10-400, institute
The flow velocity for stating oily phase is micro- l/h of 400-4000, and the microlayer model speed of production is 0.2-10 kHz.
It is further preferred that in step (1), the flow velocity of the hyperbranched poly glycerine and double mercapto-polyglycol aqueous solutions
The flow velocity of the markd protein peptides monomer solution to be captured of micro- l/h of respectively 180-220, the band be 180-220 microlitres/
Hour, the flow velocity of the oil phase is micro- l/h of 1800-2200.
One according to the present invention specific and preferred aspect, the tretolite are perfluorooctanol oil, and the cleaning uses
The phosphate buffer solution of pH 7-8 is cleaned in centrifugal rotary cup column.
According to the present invention, polyethylene glycol diamine, hyperbranched poly glycerine molecular weight can be as needed micro- cage aperture come
Design.Preferably, the molecular weight of the polyethylene glycol diamine is 1-5kDa, and the molecular weight of hyperbranched poly glycerine is 10-
30kDa.It is highly preferred that the molecular weight of the polyethylene glycol diamine is 2-3kDa, the molecular weight of hyperbranched poly glycerine is 15-
25kDa。
According to the present invention, the size of the microgel is about 30-80 microns, and preferably 40-60 is micron, more preferably
45-55 microns.
Due to the implementation of above technical scheme, the invention has the following advantages over the prior art:
Present invention innovation uses sulfydryl-alkene click-reaction combination microflow control technique method, has developed monodispersity
The micro- cage of hydrogel protein capture.Since in false folding and accumulation process, albumen grows into bigger ruler from small size monomer
Very little fiber can wash away the low small protein of folding degree according to the method for the present invention, to only capture false folding journey
High protein oligomer and fiber are spent, to carry out for Journal of Sex Research.Meanwhile the formation of hydrogel is without heating or purple
External exposure in the process will not have an impact protein structure, keep testing result more acurrate.
Detailed description of the invention
Fig. 1 is process principle figure of the invention；
Fig. 2 is the structural schematic diagram of micro-fluidic chip；
Fig. 3 shows that preparing outer layer using micro-fluidic chip is oily phase internal layer showing for the microgel containing albumen peptide solution
It is intended to；
Fig. 4 is the microlayer model of embodiment preparation and the optical microscope of micro- cage；
Fig. 5 is the fluoremetry figure of the micro- cage of hydrogel；
Fig. 6 is the microlayer model of embodiment preparation and the grain size distribution that micro- cage is observed and drawn through optical microscope；
Fig. 7 is the schematic diagram of the micro- cage thioflavine T Coloration experiment fluorescence intensity of hydrogel.
Specific embodiment
Spherical hydrogel, also known as microgel have been used for the microenvironment for simulating extracellular matrix.Under normal conditions, microgel
It can be made by refiner.But the microgel partial size that this method obtains is polydispersion, different size of microgel
Lack being compared property between body.And the comparativity between microgel sphere can be improved in the monodispersity of partial size.
Process principle figure of the invention is referring to Fig. 1.In the present invention, we have used sulfydryl-alkene click-reaction while having tied
The method for closing microflow control technique, has developed the micro- cage of hydrogel protein capture of monodispersity.Specifically, the present invention utilizes
Micro-fluidic chip production has the hydrogel microlayer model of monodispersity, and albumen peptide monomer is wrapped in microlayer model (such as Figure 1A
It is shown).Then assembled by folding protein peptides in 37 DEG C of culture microlayer models, the method for the microlayer model that is cooled with an ice bath later,
It plants albumen accumulation process (as shown in Figure 1B).Then, by the way that tretolite (such as weak surfactant) is added except deoiling and table
The drop coating of face activating agent.The addition of tretolite causes surfactant in the unstable of oil/water interface, to make grease
The separation (as shown in Figure 1 C) of two-phase.If using weak surfactant as fluorocarbon oil (such as perfluorooctanol or other contain
The fluorine compounds of one small hydrophilic radical), the protein peptides or oligomer and polymer build up body that do not assemble (are less than microgel duct
Size) it can be all washed out (as shown in figure iD) during buffer solution rinses microgel.In contrast, it is greater than microgel hole
The polyphosphazene polymer collective of road size and the protein accumulation body of threadiness are then trapped in microgel reticular structure.Meanwhile by
In the presence of microgel reticular structure, microgel can not only retain protein aggregation body, life can be added into microgel with subsequent
Small molecule needed for object reacts, to carry out other biological reaction.
The present invention will be further described in detail combined with specific embodiments below, but the present invention is not limited to following implementations
Example.The condition being not specified in embodiment is conventional laboratory conditions.
The design and production of 1 micro-fluidic chip of embodiment
Micro-fluidic chip is designed as shown in Figure 2 comprising chip body 1 is formed on chip body 1 and portion at one end
It intersects and constitutes the first microchannel 2, the second microchannel 3 and the third microchannel 4 of the first focal zone A, be formed on chip body 1
And at one end portion intersect constitute the second focal zone B the 4th microchannel 5, the 5th microchannel 6 and the 6th microchannel 7 and shape
At in the 7th microchannel 9 on chip body 1, wherein the other end of the 4th microchannel 5 is connected to the first focal zone A, the 7th is micro-
The one end in channel 9 is connected to the second focal zone B.Input end in the first to the 6th microchannel 7 is respectively arranged with filter 8.This
In example, the filtering accuracy of filter 8 is 1 micron.
Make micro-fluidic chip with Soft lithograph legal system, the specific steps are as follows:
I) designed each microchannel is printed on Acetate Film, and Acetate Film is placed in the silicon wafer coated with photoresist
At the top of piece, by ultraviolet light to get chip template；
Ii) polydimethylsiloxane prepolymer object and its crosslinking agent are poured in chip template, solidified；It will be cured
Dimethyl silicone polymer mold removes chip template, and carries out pressure piercing to the entrance of each microchannel, uses later
The substrate (such as microscope slide) of surfacing is sealed each microchannel；
Iii surface hydrophobicity agent (such as coating film on glass agent Aquapel)) is injected into microchannel and stands 5 minutes, so that channel table
Face is hydrophobic, installs filter additional finally up to micro-fluidic chip.
Embodiment 2 captures people's amyloid beta 1-42 albumen
(1) prepare outer layer be oily phase internal layer be the microgel containing albumen peptide solution
Respectively by 2- iminothiolane hydrochloride, polyethylene glycol diamine (2kDa) and hyperbranched poly glycerine (20kDa)
It is dissolved in the Napi buffer solution of pH 7.4, obtains the 2- imino group mercaptan that concentration is respectively 21g/L, 300g/L and 1000g/L
The aqueous solution of heptane hydrochloride salt, polyethylene glycol diamine and hyperbranched poly glycerine.
2- iminothiolane hydrochloric acid saline solution is mixed with polyethylene glycol diamine aqueous solution with volume ratio 0.75:1, room temperature
Lower stirring 30 minutes makes the two reaction convert mercapto-polyglycol in pairs, obtains double mercapto-polyglycol aqueous solutions, reactional equation
Formula is as follows:
It is airtight that hyperbranched poly glycerin solution and double mercapto-polyglycol aqueous solutions are injected separately into 200 microlitres of fixed pins
In syringe.
With HiLyte FluorTMThe end N- of 488 pairs of people's amyloid beta 1-42 peptide monomer side chains carries out fluorescent marker,
And it is dissolved in the ammonium hydroxide of 1wt% as preparation protein peptides monomer solution.By preparation protein peptides monomer solution with the phosphorus of pH 7.4
Acid buffering solution is diluted to required concentration (such as 5 microns), obtains HiLyte FluorTMPeople's amyloid beta 1-42 of 488 modifications
Aqueous solution (hereinafter referred to as albumen peptide solution), inject syringe, be placed in ice bath.
It is shown in Figure 3, take hyperbranched poly glycerin solution, double mercapto-polyglycol aqueous solutions, albumen peptide solution point
The first microchannel, the second microchannel and the third microchannel of micro-fluidic chip, San Zhe are not injected with 200 micro- ls/h of flow velocity
First focal zone of chip is mixed to get mixed liquor.Mixed liquor flows into the second focal zone from the 4th microchannel, and focuses second
Area is wrapped to form the monodisperse microlayer model (optics of microlayer model by the fluorination liquid HFE-7500 injected with 2000 micro- ls/h of flow velocity
Microscope figure A referring to fig. 4, for grain size distribution referring to Fig. 6 A, microlayer model size is about 48 microns, and extraordinary monodisperse is presented
Property), microlayer model is flowed out from the 7th microchannel to be collected.Generating process of the microlayer model on chip is micro- by being mounted on IX71 inversion
The monochromatic phantom V7.2 camera of mirror is imaged.Then using LabVIEW 8.2 calculate drop generation frequency, about 1.2 thousand
Hertz.During this, sulfydryl-alkene click chemistry occurs in microlayer model for hyperbranched poly glycerine and double mercapto-polyglycols
Reaction, and quickly form hydrogel.
(2) it cultivates
It is oily phase internal layer by the outer layer that step (1) is prepared is that the microgel containing albumen peptide solution is placed at 37 DEG C and trains
Certain time is supported, protein peptides is made to fold aggregation.When culture terminates, ice bath cools down 60 seconds quenching reactions.
(3) it destroys the oily phase outer layer of microgel and is cleaned
Perfluorooctanol oil is added in by the system of step (2), opens the oily phase outer layer of microlayer model, and with pH7.4's
Phosphate buffer solution cleaning 3 times in centrifugal rotary cup column (0.45 μm of aperture), and be stored in the phosphate buffer solution of pH7.4,
Obtain including the micro- cage of hydrogel (optical microscope of the micro- cage of hydrogel B referring to fig. 4, the partial size answered of high level polymeric protein
For distribution map referring to Fig. 6 B, micro- cage size is about 47 microns, and extraordinary monodispersity is presented), in case fluoremetry, corresponding glimmering
Light measurement figure is referring to Fig. 5.It can be seen from the graph that the micro- cage internal protein of hydrogel is mainly to fold the high polymeric protein (albumen of degree
Aggregation) form presence.
In addition, in order to further prove that the micro- cage internal protein of hydrogel with the presence of protein aggregation body, has carried out following experiment:
The micro- cage of hydrogel obtained by Example, is added thioflavin T (Thioflavin T) and is dyed, and excites fluorescence detection in 470nm,
As shown in fig. 7, fluorescence intensity level is significantly greater than the fluorescence intensity level of eluate in micro- cage, show the product of albumen in the micro- cage of hydrogel
Poly- degree is higher, and (after thioflavin T combination b amyloid protein, this body structure of dyestuff changes, and fluorescent value increases (amyloid egg
White accumulation degree is higher, then the amount in conjunction with thioflavin T is bigger, therefore fluorescent value is also higher).
The above embodiments merely illustrate the technical concept and features of the present invention, and its object is to allow person skilled in the art
Scholar cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to the present invention
Equivalent change or modification made by Spirit Essence, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of method using the micro- cage capture albumen of hydrogel, characterized by the following steps:
(1) make hyperbranched poly glycerin solution, double mercapto-polyglycol aqueous solutions, with markd albumen peptide monomer to be captured
Solution is mixed to get mixed liquor on micro-fluidic chip, then makes the mixed liquor, oil phase substance on the micro-fluidic chip
It mixes, acquisition microlayer model, in the microlayer model, sulfydryl-alkene occur for the hyperbranched poly glycerine and double mercapto-polyglycols
Click chemistry reacts to form hydrogel, obtain outer layer be oily phase internal layer be the microgel containing albumen peptide solution；
(2) carrying out culture to the microgel makes albumen peptide monomer therein occur to fold aggregation, obtains outer layer as oily phase internal layer
The microgel of polymeric protein containing different folding degree；
(3) tretolite is added in by the system of step (2) to destroy the oily phase outer layer and be cleaned to remove
It states micro- to get the hydrogel for capturing albumen less than the polymeric protein in the microgel duct or unpolymerized albumen in microgel
Cage, the micro- cage of hydrogel for capturing albumen can be used for subsequent detection and analysis；The tretolite is perfluorooctanol oil.
2. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
Stating micro-fluidic chip includes chip body, is formed on the chip body and portion intersects and constitutes the first focal zone at one end
First microchannel, the second microchannel and third microchannel, are formed on the chip body and portion intersects composition at one end
The 4th microchannel, the 5th microchannel and the 6th microchannel of two focal zones, and the 7th microchannel being formed on chip body,
Wherein the other end of the 4th microchannel is connected to first focal zone, one end and described second of the 7th microchannel
Focal zone connection, by hyperbranched poly glycerin solution, double mercapto-polyglycol aqueous solutions, with markd protein peptides to be captured
Solution is injected separately into first microchannel, the second microchannel and third microchannel, and three mixes in first focal zone
To the mixed liquor, the oil phase substance is injected into the 5th microchannel and the 6th microchannel respectively, mixed liquor is through the 4th
Microchannel flows to the second focal zone, mixes in the second focal zone with the oil phase substance injected out of the 5th microchannel and the 6th microchannel
Conjunction forms the microlayer model.
3. the method according to claim 2 using the micro- cage capture albumen of hydrogel, it is characterised in that: the method is also wrapped
The step of micro-fluidic chip is made with Soft lithograph method is included, which includes following several steps:
I) by designed first microchannel, the second microchannel, third microchannel, the 4th microchannel, the 5th microchannel and
Six microchannels are printed on Acetate Film, and Acetate Film is placed at the top of the silicon wafer coated with photoresist, by ultraviolet lighting
It penetrates, obtains chip template；
Ii) polydimethylsiloxane prepolymer object and crosslinking agent are poured in the chip template, solidified, it will be cured poly-
Dimethyl siloxane mold removes chip template, and carries out pressure piercing to the entrance of each microchannel, uses table later
The smooth substrate in face is sealed each microchannel;
Iii each micro channel) is injected into surface hydrophobicity agent and is stood, so that microchannel surface is hydrophobic up to the micro-fluidic core
Piece.
4. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
It states and passes through the end N- first using organic fluorescence element to albumen peptide monomer side chain with markd protein peptides monomer solution to be captured
Fluorescent marker is carried out, is then dissolved in 0.5% ~ 2% ammonium hydroxide and prepares preparation protein peptides monomer solution, finally with the slow of pH 7-8
Fliud flushing is diluted to setting concentration and obtains.
5. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
State double mercapto-polyglycol aqueous solutions pass through 2- iminothiolane hydrochloride and polyethylene glycol diamine is molten for the buffering of 7-8 in pH
Reaction obtains in liquid.
6. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
Stating albumen peptide monomer is people's amyloid beta 1-40, people amyloid beta 1-42, Protein tau or a- synapse nucleoprotein.
7. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
Stating oil phase substance is selected from one of fluorination liquid, mineral substance oil, vegetable oil and fatty acid or a variety of combinations.
8. utilizing the method for the micro- cage capture albumen of hydrogel according to claim 2 or 7, it is characterised in that: step (1)
In, the flow velocity of the hyperbranched poly glycerine and double mercapto-polyglycol aqueous solutions is micro- l/h of 10-400, described has
The flow velocity of the protein peptides monomer solution to be captured of label is micro- l/h of 10-400, and the flow velocity of the oil phase is that 400-4000 is micro-
L/h, the microlayer model speed of production is 0.2-10 kHz.
9. the method according to claim 8 using the micro- cage capture albumen of hydrogel, it is characterised in that: in step (1), institute
The flow velocity for stating hyperbranched poly glycerine and double mercapto-polyglycol aqueous solutions is micro- l/h of 180-220, described with label
Protein peptides monomer solution to be captured flow velocity be micro- l/h of 180-220, it is described oil phase flow velocity be 1800-2200 microlitres/
Hour.
10. the method according to claim 1 using the micro- cage capture albumen of hydrogel, it is characterised in that: the cleaning is adopted
With the phosphate buffer solution of pH 7-8, cleaned in centrifugal rotary cup column.
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| CN108707577B (en) * | 2018-05-25 | 2023-11-10 | 江苏汇先医药技术有限公司 | Elution method and capture method of cells or biomolecules |
| CN113272449B (en) * | 2018-10-26 | 2024-03-12 | Illumina公司 | Polymer beads are adapted for DNA treatment |
| CN109655447B (en) * | 2019-01-28 | 2022-04-08 | 广东海天创新技术有限公司 | Detection system and method for microbial enumeration |
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