CN110244038A - A kind of biomolecule flow maker reaction technology and reaction unit - Google Patents
A kind of biomolecule flow maker reaction technology and reaction unit Download PDFInfo
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- CN110244038A CN110244038A CN201910538438.8A CN201910538438A CN110244038A CN 110244038 A CN110244038 A CN 110244038A CN 201910538438 A CN201910538438 A CN 201910538438A CN 110244038 A CN110244038 A CN 110244038A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
Abstract
The invention discloses a kind of biomolecule flow maker reaction technology and reaction units, belong to field of biotechnology.Biological flow maker technology; it is characterized in that; mode is stirred to react instead of traditional by the way of flow injection reaction; make each reactant contact surface (point) reaction efficiency having the same; lab scale and large-scale production suitable for various Bioconjugation products; reaction unit includes: the first container, is used to accommodate flowable first reactive component；Second container is used to accommodate flowable second reactive component；Upstream N pass joint is respectively communicated with the first container and second container, and first reactive component and the second reactive component flow into the upstream N pass joint, and mix in the upstream N pass joint, to form reaction mixture；First temperature controlling part is used to carry out temperature control to reaction mixture, reacts reaction mixture in preset temperature range；And product reservoir, it is used to collect the Biomarker products formed after each component reaction.The present invention can control flow rate by digital pump parameter, thus the accuracy controlling biochemical reaction time, and no matter reaction scale size, can keep identical reaction to pass mild mass transfer rate, realize the indifference amplification of biomarker reaction scale.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of biomolecule flow maker reaction technology and its reaction dress
It sets, lab scale and large-scale production suitable for various Bioconjugation products.
Background technique
Bioconjugation product is widely used in field of biological detection, if gold labeling antibody is colloidal gold immunochromatographimethod product
Primary raw material, marked by magnetic bead antigen or antibody are the primary raw material of Full-automatic chemiluminescence kit, biological enzyme-labelled antigen or anti-
Body is the primary raw material of enzyme linked immunological kit, and Avidin biotin labeling large biological molecule is that signal amplifies common raw material
Deng.The characteristics of above-mentioned coupling reaction is that reaction condition is mild, it is desirable that it is that the activity stabilized of product, homogeneity are good, and if desired
Large batch of coupled product is prepared, then needs to expand the scale of production by iodine, in conventional iodine, often lab scale
The reaction condition equimultiple amplification groped can not obtain the product that activity is similar, uniformity is good, it is therefore desirable to which continuous condition is excellent
Change can just obtain active product identical as lab scale, but this optimization process consumes a large amount of manpower and material resources, improves production
Cost.If the iodine of conventional gold labeling antibody and the iodine of marked by magnetic bead antigen are usually the glass flask in large size
Middle reaction, although feeding intake equal equimultiple amplification, due to the difference of the mixing effect of size flask, the mass transfer of small flask passes Wen Su
Rate is more uniform, and the reaction effect of big flask reduces, and intermediate and edge is all variant in stirring rate and reaction temperature, causes
The coupled product of big flask is declined in activity and homogeneity.
Therefore, it is necessary to provide one kind in Bioconjugation reaction, no matter the reaction of which kind of scale can remain identical
The biomarker technology of reaction efficiency and corresponding reaction unit.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of biomarker raw material activity amplification method and device,
It can be accurately adjusted the time of biochemical reaction, it is ensured that micro and big quantitative response has identical reaction efficiency, and anti-in amplification
The middle activity for guaranteeing coupled product is answered, realizes the batch production of biomolecule coupled product.
In order to solve the above technical problems, technical solution provided by the invention is as follows:
On the one hand, a kind of biomolecule flow maker reaction technology and reaction unit are provided comprising: the first container,
It is used to accommodate flowable first reactive component；Second container is used to accommodate flowable second reactive component；Upstream N
Pass joint is respectively communicated with the first container and second container, and first reactive component and the second reactive component stream
Enter the upstream N pass joint, and mixed in the upstream N pass joint, to form reaction mixture；First temperature controlling part,
It is used to carry out temperature control to reaction mixture, reacts reaction mixture in preset temperature range；And product reservoir,
It is used to collect the Biomarker products formed after each component reaction.
Preferably, between the first container and upstream N pass joint, and/or, between second container and upstream N pass joint
Equipped with the digital pump for adjusting flow velocity.
Preferably, the biomolecule flow maker reaction unit further include: at least one supplement adjusts unit, one end
It is connected to the upstream N pass joint, the other end is connected to the product reservoir, for complete under the control of the temperature of the first temperature controlling part
At adding supplemental components in the reaction mixture after reaction.
Preferably, each supplement adjust unit include: for accommodating the adding container of supplemental components, downstream N is connected
Head and the digital pump being arranged between adding container and downstream N pass joint；And the downstream N pass joint is respectively communicated with upstream
N pass joint, adding container and product reservoir.
Preferably, biomolecule flow maker reaction unit further include: second temperature control piece is used for added with benefit
The reaction mixture for filling component carries out temperature control, keeps the reaction mixture for adding supplemental components anti-in preset temperature range
It answers.
Preferably, the supplement, which adjusts unit, several, and is serially connected.
Preferably, the flow velocity of the digital pump between the first container and upstream N pass joint is set as 10-1000 μ L/
Min, the flow velocity of the digital pump between second container and upstream N pass joint are set as 10-1000 μ L/min；The supplement adjusts single
The flow velocity of the digital pump of member is set as 10-1000 μ L/min.
On the other hand, a kind of Bioconjugation product realized using above-mentioned biomolecule flow maker reaction unit is also provided
The method of large-scale production comprising following steps:
S1, the first reactive component is accommodated in the first container, and the second reactive component is accommodated in the second container；
And the upstream N pass joint is respectively communicated with the first container and second container, adjust first reactive component and the
The flow velocity of two reactive components, so that first component is diverted into the upstream N pass joint with second group, and in the upstream N
Mixing in pass joint, to form reaction mixture liquid；
S2, starting first temperature controlling part, to be carried out to the reaction mixture formed after the upstream N pass joint
Temperature control, reacts it in preset temperature range, and the preset temperature range is 4-40 DEG C；
And S3, product reservoir is utilized to collect the biomarker head product that is formed after each component reaction.
Preferably, in the step S1 further include: one end that supplement adjusts unit is connected to the upstream N pass joint, separately
One end is connected to the product reservoir；
In the step S2 further include: the reaction mixture after completing reaction under the temperature control of the first temperature controlling part
When flowing to the downstream N pass joint of the supplement adjusting unit, so that the supplemental components in the adding container are added to reaction
In mixed liquor；
Preferably, first reactive component includes nanometer or micron order carrier material (including colloidal gold, colloidal-carbon, magnetic
Pearl, quantum dot etc.), one of large biological molecule (including antibody, antigen, biological enzyme, Streptavidin etc.) or a variety of, it is described
Second reactive component includes one of the activator that can be coupled with first reactive component, large biological molecule or a variety of.
Compared with prior art, biomolecule flow maker reaction technology and reaction unit of the invention, can basis
Actual Bioconjugation reaction type addition reduces supplementary units and corresponding pipeline, adapts to various coupled product productions with this
Needs；Bioconjugation reaction carries out in the fluid of constant temperature constant speed, and the reaction of different scales is made to all have identical pass mildly
Mass transfer rate realizes the active homogeneity of coupled product and stability；The present invention is according to gold labeling antibody and marked by magnetic bead antigen
Reaction step and feature, devise flow injection label method, Bioconjugation product prepare with scale is solved with this this
One problem realizes the active lossless amplification of various biomolecule coupled products.
Detailed description of the invention
Fig. 1 is the overall structure figure of biomolecule flow maker reaction unit in the embodiment of the present invention one；
Fig. 2 is the overall structure figure of biomolecule flow maker reaction unit in the embodiment of the present invention two；
Fig. 3 be using in embodiment two biomolecule flow maker reaction technology and its reaction unit prepare carboxyl magnetic bead
Mark the schematic diagram of Clenbuterol BSA antigen；
Fig. 4 is the overall structure figure of biomolecule flow maker reaction unit in the embodiment of the present invention three；
Fig. 5 is biomolecule flow maker reaction technology and its reaction utilized in embodiment two in the embodiment of the present invention three
Device prepares the schematic diagram of vomitoxin Immuno gold；
Fig. 6 be biomolecule flow maker reaction technology in the embodiment of the present invention three in conventional method and embodiment two and
The 25mL rank vomitoxin Immuno gold of its reaction unit preparation and the activity of lab scale (1mL) Immuno gold compare.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment one:
As shown in Figure 1, biomolecule flow maker reaction unit of the invention includes: the first container 1, being used to accommodate can
First reactive component of flowing；First reactive component includes nanometer or micron order carrier material (including colloidal gold, colloid
Carbon, magnetic bead, quantum dot etc.), one of large biological molecule (including antibody, antigen, biological enzyme, Streptavidin etc.) or more
Kind；Second container 2 is used to accommodate flowable second reactive component；Second reactive component includes can be with described first
The ingredient of reactive component coupling, specifically includes activator (including EDC/NHS, EGDE, DTT, SMCC etc.), biological macromolecule solns
(including antibody, antigen, biological enzyme, Streptavidin etc.), other solution that can be reacted or combine with first reactive component
One of or it is a variety of；
Upstream N pass joint 3, the pipeline 6 made of the low adsorptions material such as polytetrafluoroethylene (PTFE) are respectively communicated with described first
Container 1 and second container 2, and first reactive component and the second reactive component inflow upstream N pass joint 3, and
Mixing in the upstream N pass joint 3, to form reaction mixture；
First temperature controlling part 4 is used to carry out temperature control to the reaction mixture, makes it in preset temperature range
Interior reaction；Wherein, first temperature controlling part 4 includes one of water-bath component, oil bath component, refrigerator and baking oven or several
Kind, the preset temperature range is 4 DEG C -40 DEG C, selects corresponding optimum response temperature according to the condition of different kind organism coupling reaction
Degree；
And product reservoir 5, it is used to collect the Biomarker products formed after each reactive component reaction.
Preferably, the first container 1 and/or second container 2 are syringe, and the first container 1 and upstream N pass joint 3
Between, and/or, between second container 2 and upstream N pass joint 3 be equipped with adjust flow velocity digital pump 7 (including precise injection pump
Deng)；N is the integer more than or equal to 3.
Meanwhile the present embodiment additionally provides a kind of utilization above-mentioned biomolecule flow maker reaction technology and its reaction unit
The method of the Bioconjugation product prepare with scale of realization comprising following steps:
S1, flowable first reactive component is accommodated in the first container 1, and accommodating in the second container 2 can
Second reactive component of flowing；And 3 pipeline 6 made of the materials such as polytetrafluoroethylene (PTFE) of upstream N pass joint is connected respectively
Lead to the first container 1 and second container 2, first reactive component and the second reactive component are adjusted by digital pump 7
Flow velocity so that first reactive component and the second reactive component flow into the upstream N pass joint 3, and is connected in the upstream N
Mixing in first 3, to form reaction mixture；
S2, starting first temperature controlling part 4, with to the reaction mixture formed after the upstream N pass joint 3 into
Trip temperature control, reacts it in above-mentioned preset temperature range；
S3, the biomarker head product formed after each component reaction is collected using product reservoir 5.
Embodiment two:
Some Bioconjugation products prepare raw material possibly more than 2 kinds, therefore, for the system for meeting various Bioconjugation products
Standby demand, as shown in Fig. 2, the biomolecule flow maker reaction unit in the present embodiment further include: at least one supplement is adjusted
Unit 100, one end are connected to the upstream N pass joint 3, and the other end is connected to the product reservoir 5, for controlling in the first temperature
The temperature control of part 4 is lower to be completed to add supplemental components in the reaction mixture after reacting, specifically, each supplement adjusting is single
Member 100 includes: that adding container 8 (including syringe), downstream N pass joint 9 and setting are logical in adding container 8 and downstream N
Digital pump 7 (including precision digital pump) between connector 9；The supplemental components include confining liquid (including BSA, PEG, OVA, junket
Albumen, defatted milk, Tris, ethylenediamine etc.), biological macromolecule solns (including antibody, antigen, biological enzyme, Streptavidin etc.),
It can be with one in the reaction mixture reaction after completing to react under the temperature of the first temperature controlling part 4 controls or the solution combined
Kind is several.Meanwhile in the present embodiment, the biomolecule flow maker reaction unit further include: be arranged in downstream N pass joint
Between 9 and product reservoir 5 second temperature control piece 10 (including one of water-bath component, oil bath component, refrigerator and baking oven or
It is several), for adjusting the reaction mixture progress temperature control that unit 100 adds supplemental components to by supplement, mend addition
The reaction mixture for filling component reacts in above-mentioned preset temperature range, and biomolecule coupling reaction is milder, temperature range
It is 4 DEG C -40 DEG C.
It is corresponding, in the biomolecule flow maker reaction technology, the step S1 further include: supplement is adjusted into unit
One end of 100 downstream N pass joint 9 is connected to the upstream N pass joint 3, and the other end is connected to the product reservoir 5；The step
S2 further include: the reaction mixture after completing reaction under the temperature control of the first temperature controlling part 4 flows to the supplement and adjusts
When saving the downstream N pass joint 9 of unit 100, the digital pump 7 that the supplement adjusts unit 100 is opened, so that the adding container 8
Interior supplemental components are added in reaction mixture.
It is raw in the present embodiment to illustrate for preparing Clenbuterol (CLE) BSA antigenic solution of carboxyl magnetic ball label below
The structure composition and method of object molecule flow maker reaction technology and its reaction unit.
The conventional steps of Bioconjugation reaction are as follows: take 10mL 10mg/mL carboxyl magnetic ball solution (0.1M pH6.0 Mes
Make solvent), (0.1M pH5.0 Mes makees solvent to the mixed solution of addition 10mL EDC&NHS, and EDC concentration is 1.5mg/mL, NHS
Concentration is 0.75mg/mL), 37 DEG C of reaction 30min, Magneto separate is cleaned 2 times with 0.01M pH7.4 PBS, adds 10mL
0.01M pH7.4 PBS is resuspended, and 112 μ L 5.8mg/mL Clenbuterol BSA antigen stocks are added dropwise, are put in 4 DEG C of refrigerator shakings
Overnight, next day Magneto separate removes supernatant, adds magnetic bead dilution, is configured to 10mg/mL magnetic bead solution, 4 DEG C of preservations.
As shown in figure 3, in the present embodiment in biomolecule flow maker reaction unit, the first container 1, second container 2 with
And adding container 8 is syringe, upstream N pass joint 3, downstream N pass joint 9 are three-way connection, and the first temperature controlling part 4 is
Water-bath.First reactive component be 10mL 10mg/mL carboxyl magnetic ball solution (0.1M pH6.0 Mes makees solvent), second
Reactive component be 10mL EDC&NHS mixed solution (0.1M pH5.0 Mes makees solvent, wherein EDC concentration be 1.5mg/mL,
NHS concentration is 0.75mg/mL), the two is as initial reactant；The flow velocity for 2 digital pumps 7 connecting with upstream N pass joint is equal
It is set as 200 μ L/min；First temperature controlling part, 4 temperature setting is 37 DEG C；2 numbers being connect with upstream N pass joint 3
7 control of pump promotes flow velocity, mixes the first reactive component and the second reactive component at upstream N pass joint 3, forms reaction mixing
The first temperature controlling part 4 is flowed through after liquid, to complete the first step reaction within the temperature range of the first temperature controlling part 4；The benefit
It fills group and is divided into 20mL 0.033mg/mL Clenbuterol BSA antigenic solution (0.05M pH9.6 carbonate solution does dilution), with
The flow velocity for the digital pump 7 that adding container 8 is connected to is set as 400 μ L/min；Complete the reaction mixture and supplement of first step reaction
Component mixes at downstream N pass joint 9, and mixed liquor flows through second temperature control piece (being not shown in Fig. 3) and completes second step reaction,
It collects product and is put in 4 DEG C of refrigerator shakings and stay overnight, next day Magneto separate removes supernatant, adds magnetic bead dilution, is configured to 10mg/mL magnetic bead
Solution, 4 DEG C of preservations.
Further, pipeline 6 is polytetrafluoroethylene (PTFE) material, internal diameter 3mm, between upstream N pass joint 3, downstream N pass joint 9
Length of pipe be 1.7 meters, the length of pipe of downstream N pass joint 9 to collection vessel 5 is 1 meter.
Table 1 be conventional steps and through this embodiment in technology and equipment amplification preparation Clenbuterol BSA antigen magnetic
The activity comparison that ball (500mg) is reacted with lab scale (10mg), it can be seen that amplify system using the technology and equipment in the present embodiment
The activity of standby Clenbuterol BSA antigen magnetic ball is suitable with the activity of lab scale reaction, in addition promoted (each standard point it is luminous
Value is suitable or is promoted, active 100% or more accounting), and sensitivity is constant, is still 0.2ppb；And use conventional amplification anti-
The activity for the Clenbuterol BSA antigen magnetic ball that should be prepared reduces that very much (luminous value of each standard point declines, and activity is accounted for than lab scale
Than 65%-90%).
Activity (luminous value) comparison of Clenbuterol (CLE) BSA antigen product of 1 carboxyl magnetic ball of table label
Embodiment three:
The present embodiment and embodiment two the difference is that, as shown in figure 4, the biomolecule in the present embodiment flows mark
Remember in reaction unit, the supplement, which adjusts unit 100, to be had several (such as 2), and the supplement adjusts unit 100 and mutually goes here and there
Connection.It should be noted that second temperature control piece 10 in the present embodiment include water-bath component, oil bath component, refrigerator baking oven and
One or more of air-conditioning, if one of water-bath component, oil bath component, refrigerator, baking oven, then at least two concatenated benefit
It fills to adjust and is equipped with second temperature control piece 10 between unit 100, the second temperature control piece 10 is used for addition supplemental components
Reaction mixture carry out temperature control, one end connection it is upper one supplement adjust unit 100 downstream N pass joint 9, the other end company
Lead to the downstream N pass joint 9 that next supplement adjusts unit 100, and is connected to by downstream N pass joint 9 with product reservoir 5；If second
Temperature-controlling module 10 is air-conditioning, then is independently arranged, adjustable room temperature, without having connection relationship with other component.
It is corresponding, in the biomolecule flow maker reaction technology, the step S1 further include: most upstream is supplemented and is adjusted
The one end for saving the downstream N pass joint 9 of unit 100 is connected to the upstream N pass joint 3, and the other end connects next connection downstream N and connects
First 9, and the downstream N pass joint 9 for being located at most downstream is connected to the product reservoir 5；The step S2 further include: in the first temperature control
It is logical that reaction mixture after the lower completion reaction of the temperature control of product 4 flows to the downstream N that a certain supplement adjusts unit 100
When connector 9, the digital pump 7 that the supplement adjusts unit 100 is opened, so that the supplement adjusts the benefit in the adding container 8 of unit 100
Component is filled to be added in reaction mixture.
Illustrate in the present embodiment by taking the preparation of vomitoxin Immuno gold as an example below biomolecule flow maker reaction technology and
The structure composition and method of its reaction unit.
The conventional steps of Bioconjugation reaction are as follows: take 25mL K2CO3The colloidal gold solution of pH value is mixed up in 50mL round bottom
In flask, under stirring, it is added dropwise the vomitoxin antibody-solutions 1mL of 0.1mg/mL, (25 DEG C) reaction 30min of room temperature, then by
It is added dropwise to 2.5mL 10%BSA solution, after (25 DEG C) reaction 5min of room temperature, 350 μ L 2%PEG20000 solution is added dropwise, after
Continue in (25 DEG C) reaction 25min of room temperature, reaction solution is transferred in 50mL centrifuge tube, 7000rpm is centrifuged 30min, removes supernatant
Obtain vomitoxin Immuno gold stoste.
As shown in figure 5, in the present embodiment in biomolecule flow maker reaction unit, the first container 1, second container 2 with
And 2 adding containers 8 are syringe, N pass joint 3,2 downstream N pass joints 9 in upstream are three-way connection, the first temperature control
Product 4 and second temperature control assembly 10 are air-conditioning.First reactive component is to use K2CO3Mix up the colloidal gold solution of pH value
The flow velocity of the digital pump 7 of 25mL, connection the first container 1 and upstream N pass joint 3 is set as 833 μ L/min, the second reaction group
It is divided into the vomitoxin antibody-solutions 1mL of 0.1mg/mL, the flow velocity of connection second container 2 and the digital pump 7 of upstream N pass joint 3
33 μ L/min are set as, the two is as initial reactant；Room temperature is adjusted to 25 by 4 temperature of the first temperature controlling part
℃；2 digital pumps 7 control connecting with upstream N pass joint promotes flow velocity, makes the first reactive component and the second reactive component upper
It swims and is mixed at N pass joint 3, form reaction mixture, it is anti-to complete the first step within the temperature range of the first temperature controlling part 4
It answers；Supplemental components in the supplementary units 100 of most upstream are that (i.e. sealer 1 in Fig. 5 is molten for 2.5mL 10%BSA solution
Liquid), the flow velocity of the digital pump 7 of the supplementary units 100 of the most upstream is set as 83 μ L/min, and the opening time flow to for reaction solution
When the entrance of the downstream N pass joint 9 of the supplementary units 100 of most upstream, to add in the reaction mixture for completing first step reaction
Add the supplemental components (i.e. 1 solution of sealer) in the supplementary units 100 of most upstream, and makes the supplementary units added with most upstream
The mixed liquor of supplemental components (i.e. 1 solution of sealer) in 100 mixes at the downstream N pass joint 9 of the supplementary units 100 of most upstream
It closes, mixing liquid stream (i.e. 25 DEG C) in the preset temperature range of the second temperature control piece 10 of the supplementary units 100 of most upstream is complete
It is reacted at second step；Supplemental components in next supplementary units 100 are 350 μ L 2%PEG20000 solution (i.e. closings in Fig. 5
2 solution of agent), the flow velocity of the digital pump 7 of next supplementary units 100 is set as 14 μ L/min, and the opening time is reaction liquid stream
To next supplementary units 100 downstream N pass joint 9 entrance when, with complete second step reaction reaction mixture in add
Supplemental components (i.e. 2 solution of sealer) in next supplementary units 100, and make added with the benefit in next supplementary units 100
The mixed liquor for filling component (i.e. 2 solution of sealer) mixes at the downstream N pass joint 9 of next supplementary units 100, and mixing liquid stream exists
(i.e. 25 DEG C) third step is reacted in the preset temperature range of the second temperature control piece 10 of next supplementary units 100, and is produced
Object；After collection of products, 7000rpm is centrifuged 30min, removes supernatant up to vomitoxin Immuno gold stoste.
Further, pipeline is polytetrafluoroethylene (PTFE) material, internal diameter 3mm, the supplementary units of upstream N pass joint 3 to most upstream
Length of pipe between 100 downstream N pass joint 9 is 3.7 meters, between the downstream N pass joint 9 of adjacent two supplementary units 100
Length of pipe is 0.7 meter, and the length of pipe of downstream N pass joint 9 to the collection vessel 5 of the supplementary units 100 of most downstream is 3 meters.
Table 2, Fig. 6 be conventional steps and through this embodiment in technology and equipment preparation vomitoxin Immuno gold
The activity of (25mL) and lab scale (1mL) Immuno gold compares, and is tested using double parallel, and test strips are quantitative for the vomitoxin with batch
Test paper has drawn two detection lines (T1, T2) and a nature controlling line (C) respectively.Lab scale be obtained after optimum experimental it is best
Active vomitoxin Immuno gold, the coupled product of the comparable therewith amplification scale of activity in order to obtain, one is use this reality
The Immuno gold of the technology and equipment amplification preparation in example is applied, another kind is the Immuno gold of conventional method amplification preparation, it can be seen that
The Immuno gold activity of technology and equipment preparation in through this embodiment is suitable with lab scale Immuno gold activity, i.e., T1, T2 and C line are aobvious
Color is normal, and signal value is suitable, and the activity of the Immuno gold of conventional steps amplification preparation is substantially reduced, and the colour developing of T1 and T2 line is weaker,
Its signal value is significantly lower than lab scale Immuno gold.Signal value with T1, T2 and C line of readout instrument reading is as shown in table 2, conventional to amplify
Step T1 signal value decline about 50%, T2 signal value decline about 30%, and through this embodiment in technology and equipment amplification
T1 the and T2 signal value of the vomitoxin Immuno gold of preparation is suitable with lab scale, or even slightly improves, i.e. technology in the present embodiment
The problem that activity reduces when successfully solving the amplification of standard biologic labeling method with device.
The comparison of the signal value of 2 two kinds of amplification methods of table and lab scale
In conclusion the biomarker technology and its reaction unit in the present invention are in a manner of flowing reactive instead of tradition
Stirring mode, flow rate is controlled by digital pump parameter, thus the accuracy controlling biochemical reaction time, and ensure that difference
Reaction scale all have identical reaction efficiency, solve the problems, such as standard biologic labeling method amplification when activity reduce, separately
On the one hand, which can increase or decrease reaction component (such as supplementary units 100 according to specific Bioconjugation reaction type
Number etc.), it is low in cost, be particularly suitable for the coupling reaction of biological field.
It should be noted that, in this document, such as the terms "include", "comprise" or its any other variant are intended to
Non-exclusive inclusion, so that the process, method, article or equipment including a series of elements is not only wanted including those
Element, but also including other elements that are not explicitly listed, or further include for this process, method, article or equipment
Intrinsic element.In the absence of more restrictions, the element limited by sentence "including a ...", it is not excluded that
There is also other identical elements in process, method, article or equipment including the element.And above-described embodiment one to three
In technical characteristic can carry out any combination, the technical solution being composed all belongs to the scope of protection of the present invention.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (10)
1. a kind of biomolecule flow maker reaction technology and reaction unit, suitable for the mild reaction of biological field, feature
It is, comprising: the first container is used to accommodate flowable first reactive component；Second container is used to accommodate flowable
Second reactive component；Upstream N pass joint is respectively communicated with the first container and second container, and the first reaction group
Divide and flow into the upstream N pass joint with the second reactive component, and mixed in the upstream N pass joint, is mixed with forming reaction
Liquid；First temperature controlling part is used to carry out temperature control to reaction mixture, makes reaction mixture in preset temperature range
Reaction；And product reservoir, it is used to collect the Biomarker products formed after each component reaction.
2. the mild reaction of biological field as described in claim 1, which is characterized in that coupling method is mild, and reaction temperature is 4
Micron load is received in a variety of biomarkers reaction of DEG C of -40 DEG C ranges, including but not limited to colloidal gold, colloidal-carbon, magnetic bead, quantum dot etc.
The coupling reaction of body material and large biological molecule such as antibody, antigen, biological enzyme, Streptavidin etc..
3. device as described in claim 1, which is characterized in that between the first container and upstream N pass joint, and/or, the
The digital pump for adjusting flow velocity is equipped between two containers and upstream N pass joint.
4. device as described in claim 1, which is characterized in that the biomolecule flow maker reaction technology and reaction unit
Further include: at least one supplement adjusts unit, and one end is connected to the upstream N pass joint, and the other end is connected to the product reservoir,
For adding supplemental components in the reaction mixture after completing to react under the control of the temperature of the first temperature controlling part.
5. device as claimed in claim 4, which is characterized in that it includes: for accommodating supplement that each supplement, which adjusts unit,
Adding container, downstream N pass joint and the digital pump being arranged between adding container and downstream N pass joint of component；And institute
It states downstream N pass joint and is respectively communicated with upstream N pass joint, adding container and product reservoir.
6. device as claimed in claim 5, which is characterized in that biomolecule flow maker reaction technology and reaction unit also wrap
Include: second temperature control piece is used to carry out temperature control to the reaction mixture added with supplemental components, makes to add supplementation group
The reaction mixture divided reacts in preset temperature range.
7. device as claimed in claim 4, which is characterized in that the supplement, which adjusts unit, several, and is serially connected.
8. device as claimed in claim 5, which is characterized in that the digital pump between the first container and upstream N pass joint
Flow velocity be set as 10-1000 μ L/min, the flow velocity of the digital pump between second container and upstream N pass joint is set as 10-
1000μL/min；The flow velocity for the digital pump that the supplement adjusts unit is set as 10-1000 μ L/min.
9. a kind of biomarker realized using biomolecule flow maker reaction technology described in claim 4 and reaction unit is former
Expect active amplification method, which comprises the steps of:
S1, the first reactive component is accommodated in the first container, and the second reactive component is accommodated in the second container；And it will
The upstream N pass joint is respectively communicated with the first container and second container, adjusts first reactive component and second instead
The flow velocity of component is answered, so that first component is diverted into the upstream N pass joint with second group, and is connected in the upstream N
Mixing in head, to form reaction mixture liquid；
S2, starting first temperature controlling part, to carry out temperature to the reaction mixture formed after the upstream N pass joint
Control, reacts it in preset temperature range, and the preset temperature range is 4-40 DEG C；
And S3, product reservoir is utilized to collect the biomarker head product that is formed after each component reaction.
10. method as claimed in claim 9, which is characterized in that in the step S1 further include: supplement is adjusted the one of unit
End is connected to the upstream N pass joint, and the other end is connected to the product reservoir；
In the step S2 further include: the reaction mixture flowing after completing reaction under the temperature control of the first temperature controlling part
When adjusting the downstream N pass joint of unit to the supplement, so that the supplemental components in the adding container are added to reaction mixing
In liquid.
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| CN108927082A (en) * | 2018-07-03 | 2018-12-04 | 江苏省原子医学研究所 | A kind of miniflow reaction unit of aluminum fluoride label |
| CN109030810A (en) * | 2018-05-03 | 2018-12-18 | 南安市创培电子科技有限公司 | A kind of continuous preparation method of antibody (or antigen) coating microballoon |
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| CN1953802A (en) * | 2004-01-26 | 2007-04-25 | 哈佛大学 | Fluid delivery system and method |
| CN106552561A (en) * | 2015-09-28 | 2017-04-05 | 苏州英诺凯生物医药科技有限公司 | A kind of antibody（Or antigen）The continuous preparation method of coating microsphere |
| CN207703875U (en) * | 2017-12-29 | 2018-08-07 | 上海捷门生物技术有限公司 | Myoglobin assay kit production system |
| CN109030810A (en) * | 2018-05-03 | 2018-12-18 | 南安市创培电子科技有限公司 | A kind of continuous preparation method of antibody (or antigen) coating microballoon |
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