CN207941496U - A kind of device of scale synthetic proteins nano-microcapsule - Google Patents
A kind of device of scale synthetic proteins nano-microcapsule Download PDFInfo
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- CN207941496U CN207941496U CN201721673846.7U CN201721673846U CN207941496U CN 207941496 U CN207941496 U CN 207941496U CN 201721673846 U CN201721673846 U CN 201721673846U CN 207941496 U CN207941496 U CN 207941496U
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Abstract
The utility model discloses a kind of devices of scale synthetic proteins nano-microcapsule; including the first liquid sample feeding device, second liquid injector, gas pressure tank, gas-pressure controlling valve, synthesis case, sample divider; the synthesis case includes gas flowmeter, the first fluid flowmeter, second liquid flowmeter, gas sampling pipe, Liquid sample introduction pipe, liquid mixing tube and gas-liquid mixing tube; the utility model can be used for the production of multiple protein pharmaceutical carrier, have many advantages, such as operation temperature is low, material mixing evenly, output is big, method is simple, wide adaptability.
Description
Technical field
The invention belongs to protein production technology fields, more particularly, and in particular to a kind of to be advised using microtube device
The method of modelling synthetic proteins nano-microcapsule.
Background technology
The synthetic proteins nano-microcapsule method being widely used at present is mainly mechanical agitation methods, however churned mechanically body
System can deposit the problems such as stirring is uneven, temperature gradient is poor, and it is uneven that these problems can directly contribute nano-microcapsule particle diameter distribution.It is right
For expensive protein medicaments, stirring can cause local temperature excessively high, may influence the activity of protein drug.
Invention content
It is an object of the invention to overcome the deficiencies in the prior art to provide one kind for disadvantages mentioned above in the prior art
The device and production method of at low cost, easy to operate, the uniform particle sizes that are produced on a large scale protein nano micro-capsules.
The technical purpose of the present invention is achieved by following technical proposals：
Microchannel protein nano micro-capsule synthesizer, including the first liquid sample feeding device, second liquid injector, gas pressure
Tank, gas-pressure controlling valve, synthesis case, sample divider, the synthesis case include gas flowmeter, the first fluid flowmeter,
Second liquid flowmeter, gas sampling pipe, the first Liquid sample introduction pipe, second liquid sample feeding pipe, liquid mixing tube and gas-liquid mixed
Pipe forms.
Wherein：The air outlet of gas pressure tank enters gas-pressure controlling valve, the gas pressure by piping connection
The outlet of control valve is separated is divided into the output of three tunnels by gas, is respectively connected to entrance, the first liquid of gas sampling pipe in synthesis case
The gas phase entrance of injector, the gas phase entrance of second liquid injector, the outlet of the gas sampling pipe pass through pipeline and gas
The entrance of flowmeter is connected, and outlet and the first fluid flowmeter of first liquid sample feeding device pass through the first Liquid sample introduction pipe phase
Even, the outlet of the second liquid injector is connected with second liquid flowmeter by second liquid sample feeding pipe, the first liquid flow
The outlet of gauge is connect with the outlet of second liquid flowmeter by threeway converge after access liquid mixing tube, liquid mixing
The outlet of pipe and the outlet of gas flowmeter access gas-liquid mixed pipe after converging by threeway connection, product is from the gas-liquid mixed
The outlet of pipe obtains.
In above-mentioned technical proposal, the liquid mixing length of tube be 5-30cm, along all flowmeters in Flow of Goods and Materials direction it
The internal diameter of pipeline afterwards is preferably 0.5-10mm.
In above-mentioned technical proposal, the gas-liquid mixed length of tube is 10cm-150cm.
In above-mentioned technical proposal, first liquid sample feeding device and the second liquid injector should have air-tightness special
Sign.
In above-mentioned technical proposal, the outlet and the outlet of the second liquid flowmeter of first fluid flowmeter pass through
T-type threeway connection accesses the liquid mixing tube, the outlet of the liquid mixing tube and going out for the gas flowmeter after converging
Mouth accesses the gas-liquid mixed pipe after converging by Y-type three way type connection.
In above-mentioned technical proposal, the pressure-control valve is controlled by computer control terminal.
In above-mentioned technical proposal, the outlet connection sample divider of the gas-liquid mixed pipe carries out product collection.
In above-mentioned technical proposal, the pipe material after all flowmeters in Flow of Goods and Materials direction is preferably polytetrafluoroethylene (PTFE)
Material.
Scale protein nano micro-capsule synthetic method is carried out using above-mentioned processing unit, is carried out by the following method：
Using albumen, monomer, crosslinking agent, catalyst aqueous premix as the first solution, inject the first liquid sample feeding device,
Initiator is configured to buffer solution as the second solution, injects second liquid injector, the first solution and the second solution pass through
Gas-liquid mixed pipe is mixed into inert protective gas again after the mixing of Liquid sample introduction pipe, gas-liquid mixture is with drops stream
It is dynamic, it is finally exported in gas-liquid mixed pipe and obtains protein nano micro-capsule.
In the above-mentioned technical solutions, albumen selection bovine serum albumin(BSA), glucose oxidase, horseradish in first solution
Peroxidase or monoclonal antibody, monomer select 2- methylacryoyloxyethyls Phosphorylcholine (MPC), acrylic acid, metering system
Acid, acrylamide or Methacrylamide, crosslinking agent are bisacrylamide or ethylene glycol dimethacrylate, catalyst choice
Fat amine compound or aliphatic diamine class compound.
In the above-mentioned technical solutions, initiator selects persulfuric acid salt compounds in second solution.
In the above-mentioned technical solutions, one kind in inert protective gas selection nitrogen, helium, argon gas.
In the above-mentioned technical solutions, a concentration of 1mg/mL-10mg/mL of protein solution, albumen: monomer: crosslinking agent: catalyst
Molar ratio be 1: (300-10000): (300-800): (300-800).
In the above-mentioned technical solutions, the molar ratio of albumen and initiator is 1: (300-800).
In the above-mentioned technical solutions, before injecting the liquid sample feeding device, the first solution and the second solution use PB respectively
It is isometric that buffer solution is adjusted to two kinds of solution.
In above-mentioned technical proposal, the inert protective gas flow into gas-liquid mixed pipe is 50 μ L/min-8000 μ L/min,
Control makes the first liquid sample feeding device and the into the inert protective gas pressure of the first liquid sample feeding device and second liquid injector
Gas pressure is not less than the gas flowmeter outlet gas pressure in two liquid sample feeding devices.
In above-mentioned technical proposal, it is respectively 10 μ L/ to control the first solution and the second solution to enter the flow of liquid mixing tube
min-4000μL/min。
In above-mentioned technical proposal, the flow-rate ratio of first solution and second solution into the liquid mixing tube is
(0.5-2): 1, liquid inventory and gas flow ratio are 1 in the gas-liquid mixed pipe: (2-6).
In above-mentioned technical proposal, system inject raw material before using the inert protective gas to entire synthesizer into
Remaining air in row purging discharge system.
In the above-mentioned technical solutions, finally obtained product protein nano micro-capsule is to be by core, high molecular material of albumen
The nucleocapsid of shell.
Compared with prior art, the first solution to react to each other and the second solution are formed after mixing in liquid mixing duct
The mixed liquor continuously flowed, then converges at threeway with gas, and gas is by the liquid separation continuously flowed at uniform
Droplet, one side gas have shear action to liquid, promote the first solution and the mixing of the second solution, on the other hand reaction is small
It is carried out in this small system of drop, is conducive to the physical diffusion mixing of the first solution and the second solution, droplet is mixed in gas-liquid
It closes and is flowed in pipeline, polymerisation is carried out in gas-liquid mixed pipe with drops.Compared with the production technology of traditional reaction kettle,
Its design of the invention is clear, easy to operate, has the characteristics that heat-transfer effect is good, integrated, and it is uniform can to provide medium for reaction
Reaction environment is suitable for the production of multiple protein pharmaceutical carrier.
Description of the drawings
The structural schematic diagram of Fig. 1 microchannel protein nano micro-capsule synthesizers；
In figure：1. computer control terminal；2. gas-pressure controlling valve；3. gas pressure tank；4. the first liquid sample feeding device；5. the
Two liquid sample feeding devices；6. synthesizing case；7. sample divider.
The synthesis case internal structure schematic diagram of Fig. 2 microchannel protein nano micro-capsule synthesizers；
In figure：8. gas sampling pipe；9. the first Liquid sample introduction pipe；10. second liquid sample feeding pipe；11. gas flowmeter；
12. the first fluid flowmeter；13. 14. liquid mixing tube of second liquid flowmeter；15 gas-liquid mixed pipes.
The grain-size graph of 2 bovine serum albumin(BSA) nano-microcapsule of Fig. 3 embodiments；
The potential diagram of 2 bovine serum albumin(BSA) nano-microcapsule of Fig. 4 embodiments；
The transmission plot of 2 bovine serum albumin(BSA) nano-microcapsule of Fig. 5 embodiments；
The electrophoretogram of 2 bovine serum albumin(BSA) nano-microcapsule of Fig. 6 embodiments.
Specific implementation mode：
In order to make advantages of the present invention, technical solution and purpose be more clearly understood that, with reference to example to the present invention into traveling
The explanation of one step.The case study on implementation of the present invention is given below, is the further explanation to the present invention, rather than limits the present invention's
Range.
Case study on implementation 1
The air outlet of gas pressure tank enters gas-pressure controlling valve, the gas-pressure controlling valve by piping connection
Outlet separate by gas be divided into three tunnels output, be respectively connected to synthesis case in gas sampling pipe entrance, the first liquid sample feeding device
Gas phase entrance, second liquid injector gas phase entrance, the outlet of the gas sampling pipe passes through pipeline and gas flowmeter
Entrance be connected, the outlet of first liquid sample feeding device is connected with the first fluid flowmeter by the first Liquid sample introduction pipe, institute
The outlet for stating second liquid injector is connected with second liquid flowmeter by second liquid sample feeding pipe, the first fluid flowmeter
Outlet connect to converge and be followed by by the polytetrafluoroethylene (PTFE) pipeline that internal diameter is 1mm with the outlet of second liquid flowmeter with T-type threeway
Enter the polytetrafluoroethylene (PTFE) liquid mixing tube that internal diameter is 1mm, the outlet of the liquid mixing tube and the outlet of gas flowmeter pass through
The polytetrafluoroethylene (PTFE) pipeline that internal diameter is 1mm is connect with Y-type three way type converge after access internal diameter be 1mm polytetrafluoroethylene (PTFE) gas-liquid mixed
Pipe, protein nano micro-capsule product enter sample divider from the outlet of the gas-liquid mixed pipe.
Case study on implementation 2
By 10mL10mg/mL bovine serum albumin(BSA)s, 55.8mg monomer MPC, 13.4mg monomeric acrylamide, 11.6mg crosslinkings
Agent bisacrylamide, 51mg catalyst tetramethylethylenediamines are dissolved in PB buffer solutions (20mM), are made into the first solution of 100mL.
21mg ammonium persulfate initiators are dissolved in PB buffer solutions and are made into the second solution of 100mL, the first solution, the second solution are separately added into
Into two liquid sample feeding devices, it is 320 μ L/ to control nitrogen to flow into the gas flow of gas sampling pipe (8) by computer control terminal
Min, the flow that two kinds of liquid in Liquid sample introduction pipe are controlled by fluid flowmeter is 80 μ L/min, liquid mixing length of tube
For 5cm, gas-liquid mixed length of tube is 20cm, and gas-liquid mixture is flowed with drops, then carries out the collection and packing of sample.
Fig. 3,4,5,6 are respectively grain-size graph, potential diagram, transmission plot, the electrophoretogram of the protein nano micro-capsule synthesized.Nano-microcapsule grain size
Figure.By dynamic light scattering particle size instrument (90plus PLAS, Brookhaven, UK) with 90 ° of angle of scatterings to the grain size of nano-microcapsule
It is detected.From figure 3, it can be seen that the grain size of bovine serum albumin(BSA) is about 4nm, and protein nano micro-capsule major part grain size
Between 18-28nm, the increase of grain size illustrates that BSA is wrapped up by high molecular material, the successful preparation of nano-microcapsule.
Fig. 4 nano-microcapsule potential diagrams.By zeta potential instrument (90plus PLAS, Brookhaven, UK) in 25 DEG C of conditions
The lower current potential for measuring bovine serum albumin(BSA) and nano-microcapsule.As can be seen from Figure 4 the current potential of bovine serum albumin(BSA) be -15mV ±
The current potential of 3mV, nano-microcapsule are 5mV ± 2mV.Current potential is illustrated the macromolecule material on nano-microcapsule surface by the transformation of negative electricity to positive electricity
Material shields the elecrtonegativity of bovine serum albumin(BSA), so that nano-microcapsule shows electropositive.
The transmission electron microscope picture of Fig. 5 nano-microcapsules.It is observed by transmission electron microscope (JEOF JEM-2100F, Japan)
The pattern and size of nano-microcapsule.From figure 5 it can be seen that the grain size of nano-microcapsule is 20nm or so and the spherical of rule is presented
Structure.
The electrophoretogram of Fig. 6 nano-microcapsules.It after electrophoresis 10min, is used under 120V voltage conditions by DYY-6C types electrophoresis apparatus
ZF-5 ultraviolet analyzers observe the pillar location of bovine serum albumin(BSA) and nano-microcapsule at excitation wavelength 365nm.Cow's serum
Albumin as a control group, can be mobile to anode under voltage effect since bovine serum albumin(BSA) is negative electricity.And nano-microcapsule group
Band all stays in hole, this is that high molecular material is coated on the surface of BSA so that the elecrtonegativity of BSA is shielded, this just says
The successful preparation of bright nano-microcapsule.
Case study on implementation 3
By 10mL10mg/mL bovine serum albumin(BSA)s, 34mg monomer MPC, 18mg monomeric acrylamide, 5mg crosslinking agents double third
Acrylamide, 26mg catalyst tetramethylethylenediamines are dissolved in PB buffer solutions (20mM), are made into the first solution of 100m.13mg is drawn
Hair agent ammonium persulfate is dissolved in PB buffer solutions and is made into the second solution of 100mL, and the first solution, the second solution are added separately to two liquid
In body injector, it is 600 μ L/min to control nitrogen to flow into the gas flow of gas sampling pipe (8) by computer control terminal, is passed through
It is 100 μ L/min that fluid flowmeter, which controls the flow in two Liquid sample introduction pipes, and liquid mixing length of tube is 10cm, and gas-liquid is mixed
Conjunction length of tube is 60cm, and gas-liquid mixture is flowed with drops, then carries out the collection and packing of sample.
Case study on implementation 4
By 10mL10mg/mL bovine serum albumin(BSA)s, 80mg monomer MPC, 20mg monomeric acrylamide, 16mg crosslinking agents double third
Acrylamide, 32mg catalyst tetramethylethylenediamines are dissolved in PB buffer solutions (20mM), are made into the first solution of 100mL.By 20mg
Initiator ammonium persulfate is dissolved in PB buffer solutions and is made into the second solution of 100mL, and the first solution, the second solution are added separately to two
In liquid sample feeding device, it is 1500 μ L/min to control nitrogen to flow into the gas flow of gas sampling pipe (8) by computer control terminal, is led to
It is 600 μ L/min to cross the flow that fluid flowmeter controls in two Liquid sample introduction pipes, and liquid mixing length of tube is 12cm, gas-liquid
Mixing length of tube is 100cm, and gas-liquid mixture is flowed with drops, then carries out the collection and packing of sample.
Case study on implementation 5
By 10mL10mg/mL bovine serum albumin(BSA)s, 40mg monomer AAM, 20mg monomeric acrylamide, 16mg crosslinking agents double third
Acrylamide, 32mg catalyst tetramethylethylenediamines are dissolved in PB buffer solutions (20mM), are made into the first solution of 100mL.By 20mg
Initiator ammonium persulfate is dissolved in PB buffer solutions and is made into the second solution of 100mL, and the first solution, the second solution are added separately to two
In liquid sample feeding device, it is 600 μ L/min to control nitrogen to flow into the gas flow of gas sampling pipe (8) by computer control terminal, is led to
It is 100 μ L/min to cross the flow that fluid flowmeter controls in two Liquid sample introduction pipes, and liquid mixing length of tube is 10cm, gas-liquid
Mixing length of tube is 60cm, and gas-liquid mixture is flowed with drops, then carries out the collection and packing of sample.
Case study on implementation 6
10mL10mg/mL glucose oxidases, 48mg monomer MPC, 7.2mg monomeric acrylamide, 5.2mg crosslinking agents is double
Acrylamide, 12mg catalyst tetramethylethylenediamines are dissolved in PB buffer solutions (20mM), are made into the first solution of 100mL.It will
10mg initiator ammonium persulfates are dissolved in PB buffer solutions and are made into the second solution of 100mL, and the first solution, the second solution are added separately to
In two liquid sample feeding devices, it is 320 μ L/ to control nitrogen to flow into the gas flow of gas sampling pipe (8) by computer control terminal
Min, the flow that two kinds of liquid in Liquid sample introduction pipe are controlled by fluid flowmeter is 80 μ L/min, liquid mixing length of tube
For 5cm, gas-liquid mixed length of tube is 20cm, and gas-liquid mixture is flowed with drops, then carries out the collection and packing of sample.
Claims (8)
1. a kind of device of scale synthetic proteins nano-microcapsule, including the first liquid sample feeding device, second liquid injector, gas
Pressurized tank, gas-pressure controlling valve, synthesis case, the synthesis case includes gas flowmeter, the first fluid flowmeter, second liquid
Flowmeter, gas sampling pipe, the first Liquid sample introduction pipe, second liquid sample feeding pipe, liquid mixing tube and gas-liquid mixing tube composition；
Wherein：The air outlet of gas pressure tank enters gas-pressure controlling valve, the gas pressure control by piping connection
The outlet of valve is separated is divided into the output of three tunnels by gas, is respectively connected to entrance, the first Liquid sample introduction of gas sampling pipe in synthesis case
The gas phase entrance of device, the gas phase entrance of second liquid injector, the outlet of the gas sampling pipe pass through pipeline and gas flow
The entrance of meter is connected, and the outlet of first liquid sample feeding device is connected with the first fluid flowmeter by the first Liquid sample introduction pipe,
The outlet of the second liquid injector is connected with second liquid flowmeter by second liquid sample feeding pipe, the first fluid flowmeter
Outlet connect by threeway with the outlet of second liquid flowmeter converge after access liquid mixing tube, the liquid mixing tube
Outlet is connect with the outlet of gas flowmeter by threeway converge after access gas-liquid mixed pipe, product is from the gas-liquid mixed pipe
Outlet obtains.
2. a kind of device of scale synthetic proteins nano-microcapsule, the liquid mixing length of tube are according to claim 1
5-30cm, the gas-liquid mixed length of tube are 10cm-150cm.
3. a kind of device of scale synthetic proteins nano-microcapsule according to claim 1, along all streams in Flow of Goods and Materials direction
The internal diameter of pipeline after gauge is preferably 0.5-10mm.
4. a kind of device of scale synthetic proteins nano-microcapsule according to claim 1, first liquid sample feeding device and
The second liquid injector should have airtightness features.
5. a kind of device of scale synthetic proteins nano-microcapsule, the pressure-control valve pass through electricity according to claim 1
Brain control terminal is controlled.
6. a kind of device of scale synthetic proteins nano-microcapsule according to claim 1, first fluid flowmeter
Outlet is connect with the outlet of the second liquid flowmeter by T-type threeway converge after access the liquid mixing tube, the liquid
The outlet of body mixing tube is connect with the outlet of the gas flowmeter by Y-type three way type converge after access the gas-liquid mixed pipe.
7. a kind of device of scale synthetic proteins nano-microcapsule according to claim 1, the outlet of the gas-liquid mixed pipe
It connects sample divider and carries out product collection.
8. a kind of device of scale synthetic proteins nano-microcapsule according to claim 1, along all streams in Flow of Goods and Materials direction
Pipe material after gauge is preferably polytetrafluoroethylene material.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109865481A (en) * | 2017-12-05 | 2019-06-11 | 天津大学 | A kind of device and its production method of scale synthetic proteins nano-microcapsule |
| CN113975227A (en) * | 2020-07-10 | 2022-01-28 | 天津大学 | Flow type synthesis device and synthesis method of nanogel |
-
2017
- 2017-12-05 CN CN201721673846.7U patent/CN207941496U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109865481A (en) * | 2017-12-05 | 2019-06-11 | 天津大学 | A kind of device and its production method of scale synthetic proteins nano-microcapsule |
| CN113975227A (en) * | 2020-07-10 | 2022-01-28 | 天津大学 | Flow type synthesis device and synthesis method of nanogel |
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