CN107588997A - A kind of high throughput protein sample pretreatment equipment - Google Patents
A kind of high throughput protein sample pretreatment equipment Download PDFInfo
- Publication number
- CN107588997A CN107588997A CN201610537191.4A CN201610537191A CN107588997A CN 107588997 A CN107588997 A CN 107588997A CN 201610537191 A CN201610537191 A CN 201610537191A CN 107588997 A CN107588997 A CN 107588997A
- Authority
- CN
- China
- Prior art keywords
- temperature
- reductor
- denatured
- solvent
- pump
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a kind of high throughput protein sample pretreatment equipment, is a kind of collection protein denaturation and reduction, desalination and system digested online online, including：Protein processing module, power module, heating control module, pump and pump control module.It is characteristic of the invention that optimized design has been done between each operation module of whole equipment, modules are integrated into an operation device with optimal mode.Heating and control module have been designed and produced according to the requirement of flow and temperature；Small volume is selected according under the premise of meeting the resistance of device and exchanging flow quantity requirement, in light weight, the piezoelectric pump of power consumptive province；Temperature sensor is placed in optimal temperature measuring point；Whole device is simple in construction, esy to use, safe and reliable, there is preferable practicality.
Description
Technical field
The present invention relates to a kind of high throughput protein sample pretreatment equipment, pass through power module, heating control module, pump
And the control of pump control module module, complete protein example pretreatment.
Background technology
Traditional protein group sample pretreatment includes the steps such as denaturation, reduction, alkylation, desalination and enzymolysis, generally
It is offline to carry out by hand, time length is not only spent, and there is sample pollution, lose, have a strong impact on scale
The result of protein analysis.Therefore develop online sample pretreating method and device has very important significance, and at present
The equipmentization of relevant apparatus not yet has been reported that.
It is pre- that we have developed a kind of protein sample for possessing online albuminous degeneration, reduction, desalination and online enzymolysis function
Processing equipment.The equipment can realize high flux, the protein example pretreatment of low loss, have in proteomics research
Good application prospect.
The content of the invention
To integrate protein denaturation, reduction, desalination and the online sample that digests pre- it is an object of the present invention to provide a kind of
Processing unit, the realization of the function need to control it is corresponding exchange liquid conveying, control is high-temperature denatured and reduction and certain temperature
Enzyme reaction, therefore the present invention adopts the following technical scheme that.
A kind of high throughput protein sample pretreatment equipment, including：Protein pretreatment module, power module, heating control
Molding block, pump and pump control module.
Position is increased successively in a device for high-temperature denatured and reductor, solvent displacer and enzyme reactor；It is high-temperature denatured
And reductor, solvent displacer have inclination angle with horizontal plane；High-temperature denatured and reductor, solvent displacer both sides are entered using lower end
Liquid mouth, upper end liquid outlet, for ensureing that exchanging liquid is full of inside cavity, make exchange abundant；It is convenient to change, it is high-temperature denatured and
Reductor, exchange liquid pipe with their all of the above disengaging between solvent displacer and enzyme reactor and all connected with screw socket activity
Connect.
Supported outside high-temperature denatured and reductor, solvent displacer and enzyme reactor with nylon material machining block,
Play heat insulation effect simultaneously.
High-temperature denatured and reductor, temperature sensor and heat lead on solvent displacer and enzyme reactor by
USB interface is drawn to be connected respectively at temperature controller and heating power supply.
The voltage and power of high-temperature denatured and reductor and enzyme reactor power supply exchange liquid according to high-temperature denatured and reductor
It is required that the temperature reached and the flow for exchanging liquid, and the reaction temperature of enzyme reactor require conversion and obtained, while ensureing temperature
On the premise of degree requires, respectively with 36 and 24 volts of safe voltage, make the operation power utilization environment of instrument safe.
High-temperature denatured and reductor temperature sensing probe, which is directly inserted in, to be exchanged in liquid, the TEMP of enzyme reactor power supply
Probe is placed on close to temperature measuring point position, makes measurement temperature closer to actual temperature.
Pump and its control system are high-temperature denatured and reductor, and the conveying of solvent displacer exchanges liquid and controls flow, according to
The system is actually needed, and using piezoelectric micropump, small volume is in light weight, and the parameters of pump are adjusted by the control panel of pump
Claimed condition in being worked to device, parameter is locked, do not have to adjust again in work later, convenient use.
The flow of pump is the ml/min of 1 ml/min -5
The invention has the advantages that：
1, whole instrument modules are optimized, and intermodule reaches best match.
2, small volume is in light weight, power consumptive province, securely and reliably.
3, operation and part replacement are convenient, simply.
It is characteristic of the invention that optimized design has been done between each operation module of whole equipment, with optimal mode
Modules are integrated into an operation device.Heating and control module have been designed and produced according to the requirement of flow and temperature；
Small volume is selected according under the premise of meeting the resistance of device and exchanging flow quantity requirement, in light weight, the piezoelectric pump of power consumptive province；
Temperature sensor is placed in optimal temperature measuring point；Whole device is simple in construction, esy to use, safe and reliable, there is preferable practicality
Property.
Brief description of the drawings
Fig. 1：High throughput protein sample pretreatment apparatus figure；
Fig. 2：Sample pretreatment apparatus handles the mass spectrogram of transferrins, a:Active transferrins；b:After denaturation and reduction
Transferrins；c:Transferrins after desalination；d:Transferrins enzymolysis product；e:Protein in transferrins enzymolysis product
Residue detection.
Embodiment
Embodiment 1
Sample pretreatment equipment as shown in figure 1, the equipment by high-temperature denatured and reductor, solvent displacer and enzyme reaction
Device forms.In high-temperature denatured and reductor 06, solvent displacer 07, exchanging liquid and enter from inner chamber body one end, the other end comes out,
Liquid inlet and outlet is respectively provided at lower end and upper end.The hollow-fibre membrane of protein transport vehicle, passed through from inner chamber body intermediate lateral,
The material flow of the material outflow end of hollow-fibre membrane on high-temperature denatured and reductor and the hollow-fibre membrane on solvent displacer
Enter end to be connected, the material outlet of the hollow-fibre membrane on solvent displacer is connected with enzyme reactor 08 again；It is high-temperature denatured and also
The heating of prototype 06, solvent displacer 07 and enzyme reactor 08 is supplied by power supply 05, is controlled and is provided by heating controller 03, is exchanged
Liquid is supplied by infusion pump 01, the power supply of pump by pumping source 04, flow control is provided by pump discharge controller 02.
The continuity of whole sample pretreatment process is investigated using 0.1mg/mL transferrins as sample (as shown in Figure 2 a).It is first
First transferrins is pushed into high-temperature denatured and reductor with 2 μ L/min, wherein temperature is 90 DEG C, and denaturation and reducing agent are 6M salt
Sour guanidine and 50mM dithiothreitol (DTT) (as shown in Figure 2 b)；Protein example after denaturation is passed through solvent displacer, exchanges liquid and is
50mM ammonium hydrogen carbonate (pH 8.0), under forced convertion active force, the denaturant and reducing agent of high concentration are able to remove (such as Fig. 2 c
It is shown)；Final protein quality sample fixes enzyme reactor (0.25mm i.d × 50mm) by hybrid inorganic-organic matrix, in room
Temperature is lower to realize online enzymolysis, and peptide fragment caused by enzymolysis is detected (as schemed by substance assistant laser desorpted flight time mass spectrum
Shown in 2d), as can be seen from the figure myoglobins is fully converted into peptide fragment (as shown in Figure 2 e), illustrates the instrument in a large amount of albumen
There is good application value in terms of quality sample pretreatment.
Claims (6)
1. a kind of high throughput protein sample pretreatment equipment, including：Protein pretreatment module, power module, computer heating control
Module, pump and pump control module；
It is characterized in that：
In the protein processing module, the high-temperature denatured and reductor (06), solvent displacer (07) and the enzyme that are sequentially connected in series
The outermost layer of reactor (08) is high temperature resistant quartz ampoule；High-temperature denatured and reductor (06), enzyme reactor (08) inner chamber are provided with electricity
Inner sleeve is heated, solvent displacer (07) inner chamber is provided with inner sleeve；In high-temperature denatured and reductor (06), solvent displacer (07)
Exchange liquid enter from inner sleeve one end, the other end comes out；Set among high-temperature denatured and reductor (06), solvent displacer (07)
Have hollow-fibre membrane from inner sleeve among pass through；High-temperature denatured and reductor (06), solvent displacer (07) both ends are set respectively
There is the liquid inlet and outlet for exchanging liquid；
The power module is high-temperature denatured and reductor and enzyme reactor power supply (05), respectively pumping source (04), 12V, 48V
AC-AC transformer and 5V AC-DC vertoroes, by the 220V alternating currents of input be converted to required for voltage and
Current value, the power supply of heating control module and infusion pump modules is used as by connection；
The heating control module is high-temperature denatured and reductor (06) and the heating controller (03) of enzyme reactor, is
Transmit, G1 temperature controller by connection by PT100 temperature inductors are respectively high-temperature denatured and reductor
And the offer temperature display of enzyme reactor (08) and control (06)；
The infusion pump modules, it is Japanese STOL SDMP306 piezoelectric pumps, respectively high-temperature denatured and reductor (06), solvent are put
Parallel operation (07), the disengaging power for exchanging liquid is provided by connection；
The transfusion pump control module, it is Japanese STOL MPD-200A control modules, respectively high-temperature denatured and reductor (06),
The exchange liquid delivery pump of solvent displacer (07), provide flow by connection and control.
2. according to the pre-processing device described in claim 1, it is characterised in that：
High-temperature denatured and reductor (06), solvent displacer (07) and enzyme reactor (08) increase position successively in a device
(gradually increasing apart from the position of horizontal plane)；High-temperature denatured and reductor (06), solvent displacer (07) has with horizontal plane to incline
Oblique angle (tilts, in 5 degree of angles) with horizontal plane；High-temperature denatured and reductor (06), solvent displacer (07) both sides use
Lower end inlet, upper end liquid outlet, for ensureing that exchanging liquid is full of inside cavity, make exchange abundant；Convenient, the high temperature to change
Denaturation and reductor (06), between solvent displacer (07) and enzyme reactor (08) and their all of the above disengaging exchange liquid
Pipe is all flexibly connected with screw socket；
High-temperature denatured and reductor (06), solvent displacer (07) and enzyme reactor (08) outer surface are added with nylon material machinery
Work block supports, while plays heat insulation effect；
High-temperature denatured and reductor (06), solvent displacer (07) and temperature sensor on enzyme reactor (08) and heating are drawn
Line is drawn by USB interface and connected respectively at temperature controller and heating power supply.
3. according to the pre-processing device described in claim 1 or 2, it is characterised in that：
The voltage and power of high-temperature denatured and reductor and enzyme reactor power supply (05) are handed over according to high-temperature denatured and reductor (06)
The temperature that liquid requirement reaches and the flow for exchanging liquid are changed, and the reaction temperature of enzyme reactor (08) requires conversion and obtained, simultaneously
On the premise of temperature requirement is ensured, respectively with 36 and 24 volts of safe voltage, make the operation power utilization environment of instrument safe.
4. according to the pre-processing device described in claim 1 or 2, it is characterised in that：
High-temperature denatured and reductor (06) temperature sensing probe, which is directly inserted in, to be exchanged in liquid, the temperature of enzyme reactor power supply (05)
Sensing probe is placed on close to temperature measuring point position, makes measurement temperature closer to actual temperature.
5. according to the pre-processing device described in claim 1 or 2, it is characterised in that：
Pump and its control system are high-temperature denatured and reductor (06), solvent displacer (07) conveying exchanges liquid and controls flow,
According to being actually needed for the system, using piezoelectric micropump, small volume is in light weight, the control panel that the parameters of pump are passed through into pump
Claimed condition in device work is adjusted to, parameter is locked, do not have to adjust again in work later, convenient use；
The flow of pump is the ml/min of 1 ml/min -5.
6. according to the pre-processing device described in claim 1 or 2, it is characterised in that：
High-temperature denatured and reductor, solvent displacer and the enzyme reactor contacted successively,
Described high-temperature denatured and reductor includes cannulated tunica fibrosa, denaturation and reduction reaction cavity；Using hollow-fibre membrane as
Protein transport vehicle, hollow-fibre membrane be fixed on denaturation and reduction reaction cavity in, hollow-fibre membrane both ends through denaturation and
Reduction reaction cavity side wall is stretched out outside cavity, and material inlet and outlet are provided with denaturation and reduction reaction cavity；
The solvent displacer includes cannulated tunica fibrosa, buffer exchange cavity；Transmitted by protein of hollow-fibre membrane
Carrier, hollow-fibre membrane are fixed in buffer exchange cavity, and hollow-fibre membrane both ends pass through buffer exchange cavity side wall
Stretch out outside cavity, material inlet and outlet are provided with buffer exchange cavity；
The thing of the material outflow end of hollow-fibre membrane on high-temperature denatured and reductor and the hollow-fibre membrane on solvent displacer
Stream enters end and is connected；
The enzyme reactor is made up of quartz ampoule and electrical heating inner sleeve, and electrical heating inner sleeve is placed in quartz ampoule, solvent
The material outflow end of hollow-fibre membrane on displacer is connected with the material inlet of inner sleeve, product by inner sleeve material outlet
Outflow.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610537191.4A CN107588997A (en) | 2016-07-08 | 2016-07-08 | A kind of high throughput protein sample pretreatment equipment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610537191.4A CN107588997A (en) | 2016-07-08 | 2016-07-08 | A kind of high throughput protein sample pretreatment equipment |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107588997A true CN107588997A (en) | 2018-01-16 |
Family
ID=61046003
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610537191.4A Pending CN107588997A (en) | 2016-07-08 | 2016-07-08 | A kind of high throughput protein sample pretreatment equipment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107588997A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111220429A (en) * | 2018-11-23 | 2020-06-02 | 中国科学院大连化学物理研究所 | Array high-flux protein sample pretreatment device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244371A (en) * | 2008-03-28 | 2008-08-20 | 杨志刚 | Accurate solution matching device |
| 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 |
-
2016
- 2016-07-08 CN CN201610537191.4A patent/CN107588997A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244371A (en) * | 2008-03-28 | 2008-08-20 | 杨志刚 | Accurate solution matching device |
| 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 |
Non-Patent Citations (1)
| Title |
|---|
| 夏思敏等: "利用中空纤维膜接口去除 SDS 的蛋白质组学样品预处理方法", 《第十届全国生物医药色谱及相关技术学术交流会论文集》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111220429A (en) * | 2018-11-23 | 2020-06-02 | 中国科学院大连化学物理研究所 | Array high-flux protein sample pretreatment device |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104487835B (en) | Pre-processing electrophoresis integral flask, the one-piece type capillary electrophoresis of pre-treatment and the one-piece type capillary electrophoresis method of pre-treatment | |
| CN104914209A (en) | Laboratory COD automatic analysis robot | |
| CN104893974A (en) | Portable stem cell amplification culturing device capable of being online monitored dynamically in real time | |
| CN107588997A (en) | A kind of high throughput protein sample pretreatment equipment | |
| CN105889886A (en) | Steam generating system used for medium-high-voltage and high-power electrode | |
| CN204694687U (en) | Laboratory COD automatic analysis robot | |
| CN205414754U (en) | Battery pack's automatic stringer | |
| CN116173253B (en) | Multifunctional sterilizing comprehensive test bed for pilot plant test | |
| CN205720072U (en) | Total dissolved solid value detection equipment | |
| CN115125130A (en) | DNA synthesizer | |
| CN214097496U (en) | Online sample analysis device | |
| CN206580827U (en) | Gene sequencing electrophoresis tank | |
| CN210251868U (en) | Ozone decomposition test device | |
| CN210458379U (en) | Small pilot test experimental device for electrolytic cell | |
| CN212568825U (en) | Automatic liquid feeding and sample pushing device | |
| CN106699835A (en) | Full-automatic plasma processing equipment and method | |
| CN209946095U (en) | Automatic fluorine ion measuring device for coal and coal products | |
| CN208459879U (en) | A kind of water bath thermostat preparing medical material | |
| CN114518240A (en) | Online sample analysis device | |
| CN111220429A (en) | Array high-flux protein sample pretreatment device | |
| CN104460749B (en) | A kind of air chamber attemperating unit of gas analyser | |
| CN204369865U (en) | A kind of kapillary gene-amplification system | |
| CN108102906B (en) | Parallel reactor system and parallel control experiment method | |
| CN109294909A (en) | A kind of cell observation incubator | |
| CN209076592U (en) | Homogenizer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |