CN107478472A - A kind of high-temperature denatured and reductor being used in protein example pretreatment unit - Google Patents
A kind of high-temperature denatured and reductor being used in protein example pretreatment unit Download PDFInfo
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- CN107478472A CN107478472A CN201610397347.3A CN201610397347A CN107478472A CN 107478472 A CN107478472 A CN 107478472A CN 201610397347 A CN201610397347 A CN 201610397347A CN 107478472 A CN107478472 A CN 107478472A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
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- Analytical Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
The present invention relates to a kind of high-temperature denatured and reductor being used in protein example pre-processing device, the high-temperature denatured and reductor is mainly used in protein example pretreatment unit, and its application component includes:Hollow-fibre membrane, heating film, end socket, temprature control unit.Using the mode of heating, programming rate is fast, and temperature control is stable, is observed because of euphotic visible conveniently, reproducible, is adapted to batch industrialization production.
Description
Technical field
The present invention relates to a kind of high-temperature denatured and reductor being used in protein example pre-processing device, have respectively hollow
The devices such as tunica fibrosa, heating film, end socket, temperature control are assembled into protein example pretreatment unit, in proteomics
Sample pretreatment.
Background technology
Protein analysis method based on multidimensional liquid chromatography-mass spectrography (" shotgun ") has preferable reappearance,
As currently used proteome analysis technology.However, either carry out protein qualitative or carry out quantitative analysis,
Either posttranslational modification analysis, it is required for carrying out protein denaturation offline, reduction, alkylation, desalination and enzymolysis.These
The method that step all uses processed offline, time length is not only spent, and there is sample pollution, lose, seriously
It has impact on the result of scale protein analysis.Therefore develop online sample pretreating method and device have it is very important
Meaning.
In numerous protein denaturation methods, thermal denaturation is irreversible to have more wide application prospect because speed is fast.
However, traditional thermal denaturation is carried out in water-bath, can not be with other step Online integrations.Therefore, for these problems, we
A kind of heating film being used in protein example pre-processing device, fast denaturation and reduction available for protein are developed.
The content of the invention
It is an object of the present invention to provide in a kind of denaturation of protein example pretreatment high temperature, reductor and enzyme reactor
Key function device：Heating film.To realize the function of heating film, and coordinate other corresponding accessories：Need heating film
Rate of warming is fast, stable temperature control and for the ease of installation hollow-fibre membrane and monitoring course of reaction, also requires that reaction member is saturating
Light is visual, therefore the present invention adopts the following technical scheme that：
1st, a kind of high-temperature denatured and reductor being used in protein example pre-processing device, is mainly used in protein example
Pretreatment unit, including:Hollow-fibre membrane, heating film, end socket, temperature control.
2, base material is high temperature resistant quartz glass tube, both ends open, 1000 DEG C of not modifications, rapidly rises 50 degrees/min not
Explosion；The heating film of heating layer antimony and conductive layer tin is coated with quartz glass tube respectively, heating-up temperature and power can control respectively
The thickness of plated film；Conduction collar is compiled around metal ring is fixed at the soft nickel wire two of multiply respectively, is worn with screw using the soft nickel wire of multiply
Cross and tighten, multiply nickel wire is locked with heating film surface, and be in close contact with heating film, while by external power supply lead in screw
Upper compression；Outermost layer quartz glass protective case, the protective case can be that interior zone of heating is incubated, and the electricity consumption of the internal zone of heating of and can rises
Safety protection function.
3, the end socket, using PEEK materials, the requirement of sealing intensity can be met, and can makes the alkalescence in exchanging liquid
Under environment, it is not corroded；There is a hole among end socket, hollow-fibre membrane initially passes through one end end socket hole into cavity again from another end seal
The hole of head is passed, and both ends are compressed with housing pin respectively；Quartz heater and outer jointing jacket inject heater and protective case groove
In, and with water-fast sealing glue bond；The import of liquid is exchanged on the downside of center, outlet side at the over center position.
4, the temperature instruction and control, temperature sensor is placed directly in the exchange liquid for needing to detect, by sensor
Signal guides to thermoindicating controller from cavity, and by adjusting the control temperature parameter of temperature controller, control heater supplies
Electricity or power-off, control temperature.
The invention has the advantages that：
1st, programming rate is fast, heating tube euphotic visible, installs hollow-fibre membrane and observation is convenient, safe operation.
2nd, liquid good sealing effect is exchanged.
3rd, temperature control is accurate.
4th, small volume, it is power-saving, it can be mass, it is reproducible.
Using the mode of heating, programming rate is fast, and temperature control is stable, is observed because of euphotic visible conveniently, reproducible, is adapted to
Batch industrialization production.
Brief description of the drawings
Fig. 1 be protein example pre-processing device high-temperature denatured and reductor device, temperature sensor (1), quartzy base
Bottom and heating film (2), quartz glass protective case (3), conduction collar (4), hollow-fibre membrane (5), power supply (6), exchange the entering of liquid,
Go out liquid interface pipe (7), temperature instruction and controller (8)；
Fig. 2 be protein example pre-processing device high-temperature denatured and reductor device end socket figure, quartz glass protective case
Groove (11), heating heating tube groove (12), exchange liquid passes through hole (13) into and out of mouthpiece, and temperature sensor passes through hole
(14), hollow-fibre membrane passes through hole (15).
Fig. 3, transferrins matter heating film is high-temperature denatured and reducing condition under enzymolysis product mass spectral analysis figure.
Embodiment
Embodiment 1
As illustrated in fig. 1 and 2, structure of the present invention is：It is a kind of be used for protein example pretreatment unit in it is high-temperature denatured and
Reductor, including temperature sensor 1, quartz substrate and the quartz glass protective case 3 of heating film 2, conduction collar 4, hollow-fibre membrane
5, power supply 6, exchange the liquid inlet and outlet mouthpiece 7 of liquid；
The quartz substrate and heating film 2 are the hollow quartz for the left and right ends opening that outer surface is coated with conductive heating film
Pipe, as high-temperature denatured and reductor heater；The hollow-fibre membrane 5 of tubulose is placed through in hollow quartz tube, in quartzy base
Temperature sensor 1 is placed between internal face and hollow-fibre membrane 5 inside bottom and heating film 2；In the left and right ends of quartz ampoule
Heating film outer surface be fixed with conduction collar 4, heating film is conductively connected with conduction collar 4, and power-up is picked out from conduction collar
Power lead, electrical power wire are connected with power supply 6；
Quartz ampoule is placed in a left and right ends opening and had in vain in quartz glass protective case 3, in quartz glass protective case 3
Left side both ends are respectively equipped with end socket, the two end sockets openend airtight connection of left and right two with quartz glass protective case 3 respectively, two end sockets
Respectively with the openend airtight connection of left and right two of quartz ampoule；
Two openends of hollow-fibre membrane 5 are each passed through two end sockets with being connected outside high-temperature denatured and reductor；In two
Be respectively equipped with end socket exchange liquid into and out of mouthpiece 7, exchange being connected into and out of the one end of mouthpiece 7 and quartz ampoule inside for liquid
The logical, other end outside high-temperature denatured and reductor with being connected.
One side surface of the end socket is provided with ring-type quartz glass protective case groove 11, ring-type heated quarty tube groove 12,
And it is provided with its middle part and exchanges being passed through into and out of mouthpiece through hole 13, temperature sensor through hole 14, hollow-fibre membrane for liquid
Hole 15；
Heated quarty tube groove 12 is located inside quartz glass protective case groove 11, exchanges the wearing into and out of mouthpiece of liquid
Via 13, temperature sensor are located inside heating tube groove 12 through hole 14, hollow-fibre membrane through hole 15；
Exchange the equal through hole 14, hollow-fibre membrane through hole 15 into and out of mouthpiece through hole 13, temperature sensor of liquid
For through hole；Hollow-fibre membrane passes through from hollow-fibre membrane to be passed through in hole 15 and passes through temperature with end socket airtight connection, temperature sensor 1
Degree sensor is placed in quartz ampoule through hole 14, exchanges the passing through into and out of mouthpiece into and out of mouthpiece 7 from exchange liquid of liquid
Passed through in hole 13 and with end socket airtight connection.
Also include temperature instruction and controller 8, temperature instruction and controller 8 are connected by wire with by temperature sensor；
Conduction collar 4 is indicated by electrical power wire through temperature and controller 8 is connected with power supply 6.
Temperature sensor is placed directly in by the instruction of the high-temperature denatured and temperature of reductor and control, high-temperature denatured and reductor
Need in the exchange liquid that detects, temperature sensor signal is guided on temperature instruction and controller from quartzy tube chamber, passed through
The control temperature parameter of temperature instruction and controller is adjusted, controls and heating film is powered or powered off, control in quartzy tube chamber
Temperature.
Quartz ampoule base material is high temperature resistant quartz glass tube, both ends open, 1000 DEG C of not modifications, rapidly rises 50 degree mins
Clock does not burst；The heating film that heating layer antimony and conductive layer tin are formed, heating-up temperature and power energy are coated with quartz glass tube respectively
The thickness of plated film is controlled respectively；Conduction collar using the soft nickel wire of multiply compile around, metal ring is fixed at the soft nickel wire two of multiply respectively,
With screw through tightening, multiply nickel wire is locked with heating film surface, and be in close contact with heating film, while external power supply is drawn
Line compresses on screw；
Outermost layer is quartz glass protective case, and the protective case can be that interior zone of heating is incubated, the use of the internal zone of heating of and can
Electricity plays safety protection function.
The end socket, using PEEK materials, the requirement of sealing intensity can be met, and can makes the alkalescence in exchanging liquid
Under environment, it is not corroded；There is a hole among end socket, hollow-fibre membrane initially passes through one end end socket hole into cavity again from another end seal
The hole of head is passed, and both ends are compressed with housing pin respectively；Quartz ampoule and quartz glass protective case inject heated quarty tube groove with
In quartz glass protective case groove, and with water-fast sealing glue bond；Inlet tube one end of exchange liquid in quartz ampoule is in stone
The downside of English tube hub axial location, outlet one end for exchanging liquid in quartz ampoule are in the upside of axial centerline.
Using the protein denaturation and reductor for being coated with heating film, transferrins matter at 90 DEG C with concentrated hydrochloric acid guanidine and
High-temperature denatured and reduction is realized in second-rate threitol reaction, and the protein after denaturation is digested into polypeptide through desalination carries out mass spectral analysis,
As shown in Figure 3, the results showed that, transferrins matter is preferably digested, and protein sequence coverage rate is 60% ± 3% (n=
3), illustrate that protein has obtained abundant denaturation and reduction under heating film high temperature heating conditions.
Claims (6)
1. a kind of high-temperature denatured and reductor being used in protein example pretreatment unit, including temperature sensor (1), quartz
Substrate and heating film (2), quartz glass protective case (3), conduction collar (4), hollow-fibre membrane (5), power supply (6), exchange liquid
Liquid inlet and outlet mouthpiece (7)；
It is characterized in that：
The quartz substrate and heating film (2) are the hollow quartz for the left and right ends opening that outer surface is coated with electrically conductive heating film
Pipe, as high-temperature denatured and reductor heater；The hollow-fibre membrane (5) of tubulose is placed through in hollow quartz tube, in quartz
Temperature sensor (1) is placed between the internal face and hollow-fibre membrane (5) of substrate and heating film (2) inside；In quartz ampoule
The heating film outer surface of left and right ends is fixed with conduction collar (4), and heating film is closely connected with conduction collar (4), from conductive sleeve
Electrical power wire is picked out on ring, electrical power wire is connected with power supply (6)；
Quartz ampoule is placed in the hollow quartz glass protective case (3) of a left and right ends opening, in quartz glass protective case (3)
A left side, right both ends are respectively equipped with end socket, the two end sockets openend airtight connection of left and right two with quartz glass protective case (3) respectively, and two
The end socket openend airtight connection of left and right two with quartz ampoule respectively；
Two openends of hollow-fibre membrane (5) are each passed through two end sockets with being connected outside high-temperature denatured and reductor；In two envelopes
Be respectively equipped with head exchange liquid into and out of mouthpiece (7), exchange being connected into and out of mouthpiece (7) one end and quartz ampoule inside for liquid
The logical, other end outside high-temperature denatured and reductor with being connected.
2. according to the high-temperature denatured and reductor described in claim 1, it is characterised in that：
One side surface of the end socket is provided with ring-type quartz glass protective case groove (11), ring-type heated quarty tube groove (12),
And pass through hole (14), hollow-fibre membrane into and out of mouthpiece through hole (13), temperature sensor provided with exchange liquid in its middle part
Through hole (15)；
Heated quarty tube groove (12) is located at quartz glass protective case groove (11) inside, exchanges the wearing into and out of mouthpiece of liquid
Via (13), temperature sensor are located at heating tube groove (12) inside through hole (14), hollow-fibre membrane through hole (15)；
Exchange liquid passes through hole (15) into and out of mouthpiece through hole (13), temperature sensor through hole (14), hollow-fibre membrane
It is through hole；Hollow-fibre membrane passes through from hollow-fibre membrane and passes through in hole (15) and worn with end socket airtight connection, temperature sensor
Excess temperature sensor is placed in quartz ampoule through hole (14), exchange liquid into and out of mouthpiece (7) from exchange liquid into and out of interface
Pipe pass through hole (13) in pass through and with end socket airtight connection.
3. according to the high-temperature denatured and reductor described in claim 1, it is characterised in that：Also include temperature instruction and controller
(8), temperature instruction and controller (8) are connected by wire with by temperature sensor；
Conduction collar (4) by electrical power wire through temperature indicate and controller (8) be connected with power supply (6) or temperature indicate and
Controller (8) is connected with power supply (6).
4. according to the high-temperature denatured and reductor described in claim 1, it is characterised in that：Quartz ampoule base material is high temperature resistant stone
English glass tube, both ends open, 1000 DEG C of not modifications, rapidly rise 50 degrees/min and do not burst；Heating is coated with quartz glass tube respectively
The heating film that layer antimony and conductive layer tin are formed, heating-up temperature and power can be determined by the thickness of plated film；Conduction collar uses multiply
Soft nickel wire is compiled around metal ring is fixed at the soft nickel wire two of multiply respectively, with screw through tightening, makes multiply nickel wire and heating film table
Surface cover is tight, and is in close contact with heating film, while external power supply lead is compressed on screw；
Outermost layer is quartz glass protective case, and the protective case can be that interior zone of heating is incubated, and the electricity consumption of the internal zone of heating of and can rises
Safety protection function.
5. according to the high-temperature denatured and reductor described in claim 1, it is characterised in that：The end socket, using PEEK materials, i.e.,
The requirement of sealing intensity can be met, and can makes under the alkaline environment of exchanging liquid, is not corroded；There is a hole among end socket, in
Empty fiber membrane initially passes through one end end socket hole and passed again from the hole of other end end socket into cavity, and both ends are respectively with housing pin pressure
Tightly；Quartz ampoule and quartz glass protective case inject heated quarty tube groove with quartz glass protective case groove, and with resistance to watertight
Sealing bonds；In quartz ampoule exchange liquid inlet tube one end in the downside of quartz ampoule axial centerline, positioned at quartz
Outlet one end of exchange liquid in pipe is in the upside of axial centerline.
6. according to the high-temperature denatured and reductor described in claim 3, it is characterised in that：High-temperature denatured and reductor temperature refers to
Show and control, temperature sensor is placed directly in the exchange liquid for needing to detect by high-temperature denatured and reductor, by temperature sensor
Signal is guided to from quartzy tube chamber on temperature instruction and controller, is joined by the control temperature for adjusting temperature instruction and controller
Number, control and heating film is powered or powered off, control the temperature in quartzy tube chamber.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610397347.3A CN107478472B (en) | 2016-06-07 | 2016-06-07 | High-temperature denaturation and reduction device for protein sample pretreatment device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610397347.3A CN107478472B (en) | 2016-06-07 | 2016-06-07 | High-temperature denaturation and reduction device for protein sample pretreatment device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107478472A true CN107478472A (en) | 2017-12-15 |
| CN107478472B CN107478472B (en) | 2021-01-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610397347.3A Active CN107478472B (en) | 2016-06-07 | 2016-06-07 | High-temperature denaturation and reduction device for protein sample pretreatment device |
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Citations (6)
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|---|---|---|---|---|
| CN201375878Y (en) * | 2008-11-18 | 2010-01-06 | 北京大井易通科技发展有限公司 | Hollow fiber membrane component |
| CN102243151A (en) * | 2010-09-20 | 2011-11-16 | 华东理工大学 | Zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device |
| CN103665098A (en) * | 2012-09-20 | 2014-03-26 | 中国科学院大连化学物理研究所 | Diphasic column membrane protein microreactor and application thereof |
| CN103852527A (en) * | 2012-12-05 | 2014-06-11 | 中国科学院大连化学物理研究所 | High-flux protein sample pre-treatment device |
| CN104075931A (en) * | 2013-03-29 | 2014-10-01 | 中国科学院大连化学物理研究所 | In-situ rapid preprocessing method for protein samples |
| CN203990316U (en) * | 2014-08-08 | 2014-12-10 | 中建安装工程有限公司 | A kind of counterflow exchange distillation device |
-
2016
- 2016-06-07 CN CN201610397347.3A patent/CN107478472B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201375878Y (en) * | 2008-11-18 | 2010-01-06 | 北京大井易通科技发展有限公司 | Hollow fiber membrane component |
| CN102243151A (en) * | 2010-09-20 | 2011-11-16 | 华东理工大学 | Zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device |
| CN103665098A (en) * | 2012-09-20 | 2014-03-26 | 中国科学院大连化学物理研究所 | Diphasic column membrane protein microreactor and application thereof |
| CN103852527A (en) * | 2012-12-05 | 2014-06-11 | 中国科学院大连化学物理研究所 | High-flux protein sample pre-treatment device |
| CN104075931A (en) * | 2013-03-29 | 2014-10-01 | 中国科学院大连化学物理研究所 | In-situ rapid preprocessing method for protein samples |
| CN203990316U (en) * | 2014-08-08 | 2014-12-10 | 中建安装工程有限公司 | A kind of counterflow exchange distillation device |
Non-Patent Citations (1)
| Title |
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| 高启君等: "汽-液-固三相流化床蒸发器浓缩食品液体的实验研究 ", 《食品与机械》 * |
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| CN107478472B (en) | 2021-01-26 |
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