CN218727344U - Automated protein labeling apparatus - Google Patents

Abstract

The utility model discloses an automatic protein marking device, which comprises a stirring reaction part, a reaction container and a sealing cover, wherein the reaction container is arranged on a constant-temperature magnetic stirrer, and the sealing cover is adaptive to and buckled on an opening of the reaction container; the sample adding parts are multiple groups with the same structure, and each group of sample adding parts is provided with a liquid storage container, a connecting pipeline for communicating the liquid storage container and the reaction container, and a liquid injection pump; the circulating and filtering part is provided with a hollow fiber column which is circularly communicated with the reaction vessel through a circulating pipeline, and the circulating pipeline is provided with a circulating pump and a washing and filtering unit. The utility model has the advantages that the automation degree of protein marking is improved, and the protein marking efficiency is greatly improved; the consistency of the product is greatly improved when protein marking is carried out, and the quality of the protein marked product can be ensured; the device has complete functions and convenient disassembly, simplifies and standardizes the operation flow of protein labeling reaction, and greatly improves the reaction quality.

Description

Automated protein labeling apparatus

Technical Field

The utility model belongs to the technical field of the protein mark technique and specifically relates to an automatic change protein marking device.

Background

The labeling technology is a technology for labeling a plurality of easily-measured and high-sensitivity substances on a specific antibody or protein, and the biological properties and the content of the antigen or antibody in a reaction system are detected through the enhanced amplification effect of the labels. The marker is mainly used for positioning analysis, can also carry out quantitative analysis on a sample mixed with other molecules under certain conditions, is an ideal, rapid and cheap quantitative determination method, and has wide application in molecular biology, immunohistochemistry and medical pathology.

The main purpose of protein labeling is to detect bioengineering, auxiliary detection (e.g., reliable quantification of compounds, specific detection of protein modifications) or proteins and their binding partners, which can help us to study problems of protein abundance, interactions, post-translational modifications, functions, etc. of interest.

The labeling of proteins can improve detection sensitivity and simplify the detection workflow. The chemical reactions required for labeling need to be carried out at a temperature (e.g., 4 ℃ or 25 ℃, etc.), for smaller doses of labeling reactions the reaction products are typically injected into capped vials followed by a water bath to ensure constant temperature; for the marking reaction with large dosage, the manual liquid adding is time-consuming and tedious due to more types of liquid to be added; in addition, because both the labeling reaction and the liquid adding have time requirements, the manual liquid adding is difficult to accurately control the time, and the phenomena of different reaction conditions, more side reactions of products, different product purity and the like in each bottle are easily caused.

SUMMERY OF THE UTILITY MODEL

The utility model aims to provide an automatic change protein marking device.

In order to achieve the above purpose, the utility model can adopt the following technical proposal:

automatic change protein marking device, include:

the stirring reaction part consists of a constant-temperature magnetic stirrer, a reaction container arranged on the constant-temperature magnetic stirrer and a sealing cover which is matched and buckled on an opening of the reaction container;

the sample adding parts are a plurality of groups with the same structure, and each group of the sample adding parts is provided with a liquid storage container for storing a sample, a connecting pipeline for communicating the liquid storage container with the reaction container, and a liquid injection pump for conveying sample liquid into the reaction container; and

and the circulating and filtering part is provided with a hollow fiber column for washing and filtering the sample liquid in the reaction vessel, the hollow fiber column is circularly communicated with the reaction vessel through a circulating pipeline, and the circulating pipeline is provided with a circulating pump and a washing and filtering unit.

The method has the advantages that automation of each liquid inlet and outlet pipeline, temperature control, stirring, detection and hollow fiber column washing is realized, so that the automation degree of protein labeling is improved, and the protein labeling efficiency is greatly improved; compared with the existing manual operation which is easy to generate errors, time difference and misoperation, the consistency of the product is greatly improved when the protein marking is carried out by the equipment, and the quality of the protein marked product can be ensured; in addition, the device has complete functions, is convenient to disassemble, simplifies and standardizes the operation flow of protein labeling reaction, and greatly improves the reaction quality.

Further, the circulation pipeline include with the inlet pipe way that the lower port of hollow fiber post is linked together and with the drain pipe way that the upper port of hollow fiber post is linked together, the inlet pipe way the feed liquor end with the play liquid end of play liquid pipeline all communicate in reaction vessel's inner chamber, and then form the circulation, make the circulation filtration that the reaction liquid in the reaction vessel can last.

Further, the washing and filtering unit is communicated with the liquid inlet pipeline close to the liquid outlet end, and the circulating pump is positioned on the liquid inlet pipeline between the washing and filtering unit and the hollow fiber column; specifically, the washing and filtering unit comprises:

the first three-position three-way valve is arranged on the liquid inlet pipeline;

the second three-position three-way valve is connected with the first three-position three-way valve;

the three-position five-way valve is connected with the second three-position three-way valve;

the first filter washing container is communicated with the three-position five-way valve through a first filter washing pipeline;

the second filter washing container is communicated with the three-position five-way valve through a second filter washing pipeline; and

and the third filter washing container is communicated with the three-position five-way valve through a third filter washing pipeline.

The first washing and filtering container, the second washing and filtering container and the third washing and filtering container are used for respectively containing sodium hydroxide solution, purified water and acetic acid (or PBS buffer solution), so that the circulation pipelines which are communicated with the hollow fiber column and the reaction container are matched, and the washing and filtering operation is automatically finished.

Furthermore, a liquid collecting pipeline is communicated with a side wall opening of the hollow fiber column, and a liquid outlet end of the liquid collecting pipeline is communicated with a collecting container for collecting waste liquid filtered by the hollow fiber column.

Further, a liquid outlet pipeline is connected to the liquid outlet pipeline through a third three-position three-way valve, the liquid outlet end of the liquid outlet pipeline is communicated with the collecting container, and a bubble detection device is arranged on the liquid outlet pipeline between the third three-position three-way valve and the reaction container; the bubble detection device is used for detecting the content of bubbles in the sample liquid after the hollow fiber column washing and filtering, and the washing and filtering volume is easily judged through the appearance duration of continuous bubbles in the bubble detection device.

Furthermore, the liquid inlet end of the liquid inlet pipeline is a hard straight pipe which vertically penetrates through the sealing cover and is inserted into the reaction vessel, an electric lifting device for driving the hard straight pipe to vertically move up and down is arranged on the hard straight pipe positioned outside the sealing cover, and a liquid level sensor which is flush with the lower port of the hard straight pipe positioned in the reaction vessel is arranged on the hard straight pipe; the signal output end of the liquid level sensor is electrically connected with the signal input end of a main controller of the automatic protein marking equipment, and the control output end of the main controller is electrically connected with the control input end of the electric lifting device, so that the main controller receives a liquid level signal of the liquid level sensor and controls the electric lifting device to work, and the height adjustment of the hard straight pipe at the liquid inlet end of the liquid inlet pipeline is automatically completed.

Furthermore, the sample adding parts are seven groups, and the liquid outlet end of the connecting pipeline of each group of sample adding parts penetrates through the sealing cover to extend into the inner cavity of the reaction container, so that the sealing performance is ensured, and simultaneously, various reagents can be conveniently added in the protein labeling process.

Furthermore, a PH meter is arranged in the reaction container and used for detecting the PH value of the reaction liquid in the reaction container in real time.

Furthermore, an electronic scale is arranged at the bottom of the constant-temperature magnetic stirrer and can weigh the weight of the reaction sample contained in the reaction container in real time.

Drawings

Fig. 1 is a schematic structural diagram of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

It should be noted that all the directional indicators (such as up, down, left, right, front, back, 8230; \8230;) in the embodiments of the present invention are only used to explain the relative position relationship between the components, the motion situation, etc. in a specific posture (as shown in the attached drawings), and if the specific posture is changed, the directional indicator is changed accordingly.

Furthermore, descriptions in the present application as to "first," "second," etc. are for descriptive purposes only and are not to be construed as indicating or implying relative importance or to implicitly indicate the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature.

As shown in FIG. 1, the automatic protein labeling device of the present invention comprises a stirring reaction part, a sample adding part and a circulating filtration part.

Specifically, the stirring reaction part consists of a constant-temperature magnetic stirrer 1, a reaction container 2 arranged on the constant-temperature magnetic stirrer 1, and a sealing cover 3 which is adapted to be sealed and buckled on an opening of the reaction container 2; a PH meter 4 is arranged in the reaction container 2, the lower end of the PH meter 4 extends to the position close to the bottom of the inner cavity of the reaction container 2 and is used for detecting the PH value of the reaction liquid in the reaction container 2 in real time; in addition, in order to weigh the reaction liquid in the reaction vessel 2, an electronic scale 5 may be disposed at the bottom of the constant temperature magnetic stirrer 1, and the electronic scale 5 measures the weight of the constant temperature magnetic stirrer 1, the sealing cover 3, the reaction vessel 2 and the reaction liquid therein in real time, so that the weight of the constant temperature magnetic stirrer 1, the sealing cover 3 and the reaction vessel 2 is directly deducted, and the weight of the reaction liquid is directly obtained.

The sample adding parts are multiple groups with the same structure, and each group of sample adding parts is provided with a liquid storage container for storing samples, a connecting pipeline for communicating the liquid storage container with the reaction container 2, and a liquid injection pump for conveying sample liquid into the reaction container 2. Preferably, the liquid injection pump is a peristaltic pump, the connecting pipelines are flexible pipelines, the liquid outlet end of each connecting pipeline penetrates through the sealing cover 3 and extends into the inner cavity of the reaction container 2, and the liquid inlet end of each connecting pipeline is extended to the position, close to the bottom, of the inner cavity of the liquid storage container. Specifically, the sample addition parts can be generally provided with seven groups, namely a first sample addition part 6, a second sample addition part 7, a third sample addition part 8, a fourth sample addition part 9, a fifth sample addition part 10, a sixth sample addition part 11 and a seventh sample addition part 12, wherein the first sample addition part 6 corresponds to a first liquid storage container 6.1, a first connecting pipeline 6.2 and a first liquid injection pump 6.3, the second sample addition part 7 corresponds to a second liquid storage container 7.1, a second connecting pipeline 7.2 and a second liquid injection pump 7.3, the third sample addition part 8 corresponds to a third liquid storage container 8.1, a third connecting pipeline 8.2 and a third liquid injection pump 8.3, the fourth sample addition part 9 corresponds to a fourth liquid storage container 9.1, a fourth connecting pipeline 9.2 and a fourth liquid injection pump 9.3, the fifth sample addition part 10 corresponds to a fifth liquid storage container 10.1, a fifth connecting pipeline 10.2 and a fifth liquid storage container 10.3, the sixth liquid storage container 11.2 corresponds to a sixth liquid injection pump 10.3, the seventh liquid injection pump 11.1 corresponds to a sixth liquid injection pump 12, the seventh liquid injection pump 12.1 and the seventh liquid injection pump 12.3; therefore, the first liquid storage container 6.1, the second liquid storage container 7.1, the third liquid storage container 8.1, the fourth liquid storage container 9.1, the fifth liquid storage container 10.1, the sixth liquid storage container 11.1 and the seventh liquid storage container 12.1 can respectively contain different reagents, and when the sealing performance is ensured, the reagents can be added into the reaction container 2 without mutual influence in the protein labeling process, so that the protein labeling is completed.

The circulating filter part is provided with a hollow fiber column 13 for washing and filtering the sample liquid in the reaction vessel 2, the hollow fiber column 13 is circularly communicated with the reaction vessel 2 through a circulating pipeline 14, and the circulating pipeline 14 is provided with a circulating pump 15 and a washing and filtering unit. Specifically, the circulation pipeline 14 includes a liquid inlet pipeline 14.1 communicated with the lower port of the hollow fiber column 13 (i.e., the liquid inlet of the inner cavity of the hollow fiber column 13) and a liquid outlet pipeline 14.2 communicated with the upper port of the hollow fiber column 13 (i.e., the liquid outlet of the inner cavity of the hollow fiber column 13), the liquid inlet end of the liquid inlet pipeline 14.1 and the liquid outlet end of the liquid outlet pipeline 14.2 are both communicated with the inner cavity of the reaction vessel 2, so as to form circulation, so that the reaction liquid in the reaction vessel 1 can continuously circulate through the inner cavity of the hollow fiber column 13, thereby forming continuous circulation filtration; the washing and filtering unit is communicated with the liquid inlet pipeline 14.1 close to the liquid outlet end, and the circulating pump 15 is a peristaltic pump and is positioned on the liquid inlet pipeline 14.1 between the washing and filtering unit and the hollow fiber column 13.

The filter washing unit comprises a first three-position three-way valve 16 arranged on the liquid inlet pipeline 14.1, a second three-position three-way valve 17 connected with the first three-position three-way valve 16, a three-position five-way valve 18 connected with the second three-position three-way valve 17, and a first filter washing container 22, a second filter washing container 23 and a third filter washing container 24 which are communicated with the three-position five-way valve 18 through a first filter washing pipeline 19, a second filter washing pipeline 20 and a third filter washing pipeline 21 in a one-to-one correspondence manner; the first three-position three-way valve 16, the second three-position three-way valve 17 and the three-position five-way valve 18 are all electric control valves, and are respectively filled with washing and filtering reagents through the first washing and filtering container 22, the second washing and filtering container 23 and the third washing and filtering container 24, so that the washing and filtering operations on the hollow fiber column 13 are automatically completed by matching with the circulating pipeline 14 for communicating the hollow fiber column 13 and the reaction container 2.

Further, a liquid collecting pipeline 25 is communicated with a side wall port of the hollow fiber column 13 (i.e. the outer cavity liquid outlet of the hollow fiber column 13), and a liquid outlet end of the liquid collecting pipeline 25 is communicated with a collecting container 26 for collecting the waste liquid filtered by the hollow fiber column 13.

Furthermore, a liquid outlet pipe 28 is connected to the liquid outlet pipe 14.2 of the circulation pipe 14 through a third three-position three-way valve 27, and the third three-position three-way valve 27 is also an electric control valve and ensures that the liquid outlet end of the liquid outlet pipe 28 is communicated with the inner cavity of the collection container 26. In addition, a bubble detection device 29 is arranged on the liquid outlet pipe 14.2 between the third three-position three-way valve 27 and the reaction container 2; the bubble detecting means 29 may be a bubble detecting sensor for detecting bubbles in the reaction solution after the hollow fiber column 13 is washed and filtered, and the volume of the washing and filtering is determined by the occurrence duration of the continuous bubbles in the bubble detecting means 29.

Furthermore, the liquid inlet end of the liquid inlet pipeline 14.1 of the circulating pipeline 14 can be set to vertically penetrate through the sealing cover 3 and be inserted into the hard straight pipe 30 in the reaction vessel 2, the electric lifting device 31 for driving the hard straight pipe to vertically move up and down is arranged on the hard straight pipe 30 positioned outside the sealing cover 3, and the electric lifting device 31 can be specifically an electric hydraulic cylinder or an electric telescopic rod, so that the hard straight pipe 30 is driven to vertically lift up and down by the telescopic end of the electric hydraulic cylinder or the electric telescopic rod. In addition, a liquid level sensor 32 flush with the lower port of the rigid straight pipe 30 in the reaction vessel 2 is also arranged, and the liquid level information in the reaction vessel 2 is monitored in real time through the liquid level sensor 32.

Of course, for convenient operation, the automatic protein labeling apparatus should further comprise a main controller (i.e. a single chip), the signal output terminal of the liquid level sensor 32 should be electrically connected to the signal input terminal of the main controller, and the control output terminal of the main controller should be electrically connected to the control input terminal of the electric lifting device 31, so that the main controller receives the liquid level signal and controls the electric lifting device 31 to drive the hard straight tube 30 to ascend or descend, and adjust the height of the liquid inlet end of the hard straight tube 30 in the reaction vessel 2. Meanwhile, the control input ends of the first liquid injection pump 6.3, the second liquid injection pump 7.3, the third liquid injection pump 8.3, the fourth liquid injection pump 9.3, the fifth liquid injection pump 10.3, the sixth liquid injection pump 11.3, the seventh liquid injection pump 12.3 and the circulating pump 15 are respectively and electrically connected with the control output end of the main controller, and the main controller controls the automatic opening or closing of the liquid injection pumps so as to complete automatic sample injection and automatic circulating washing and filtering of the hollow fiber column 13; the control input ends of the first three-position three-way valve 16, the second three-position three-way valve 17, the three-position five-way valve 18 and the third three-position three-way valve 27 are electrically connected with the control output end of the main controller, and the main controller controls the passage conduction of the first three-position three-way valve 16, the second three-position three-way valve 17, the three-position five-way valve 18 and the third three-position three-way valve 27; the signal output end of the PH meter 4 is electrically connected with the signal input end of the main controller, and the PH value of the reaction liquid in the reaction container 2 is transmitted to the main controller in real time; the signal output end of the electronic scale 5 is electrically connected with the signal input end of the main controller, and the weight of the reaction liquid in the reaction container 2 is transmitted to the main controller in real time; in addition, the control input end of the thermostatic magnetic stirrer 1 should also be electrically connected with the control output end of the main controller, so that the main controller controls the thermostatic magnetic stirrer 1 to work or stop.

When in use, firstly, seven reagents of prepared sodium periodate solution, ethylene glycol solution, acetic acid solution, hydroformylation enzyme solution, 0.2mol/L bicarbonate buffer solution, naBH4 (sodium borohydride) and 0.067mol/L PBS buffer solution are respectively filled into a first liquid storage container 6.1, a second liquid storage container 7.1, a third liquid storage container 8.1, a fourth liquid storage container 9.1, a fifth liquid storage container 10.1, a sixth liquid storage container 11.1 and a seventh liquid storage container 12.1, namely, the first liquid storage container 6.1 is filled with the sodium periodate solution, the second liquid storage container 7.1 is filled with the ethylene glycol solution, the third liquid storage container 8.1 is filled with the acetic acid solution, the fourth liquid storage container 9.1 is filled with the hydroformylation enzyme solution, the fifth liquid storage container 10.1 is filled with the 0.2mol/L bicarbonate buffer solution, the sixth liquid storage container 11.1 is filled with the NaBH4 (sodium borohydride) and the seventh liquid storage container 12.061 is filled with the PBS buffer solution; then liquid inlet ends of the first connecting pipeline 6.2, the second connecting pipeline 7.2, the third connecting pipeline 8.2, the fourth connecting pipeline 9.2, the fifth connecting pipeline 10.2, the sixth connecting pipeline 11.2 and the seventh connecting pipeline 12.2 respectively extend into inner cavities of the corresponding first liquid storage container 6.1, the second liquid storage container 7.1, the third liquid storage container 8.1, the fourth liquid storage container 9.1, the fifth liquid storage container 10.1, the sixth liquid storage container 11.1 and the seventh liquid storage container 12.1, which are close to the bottom, and liquid outlet ends are firstly connected by a waste liquid bottle; and then the first liquid injection pump 6.3, the second liquid injection pump 7.3, the third liquid injection pump 8.3, the fourth liquid injection pump 9.3, the fifth liquid injection pump 10.3, the sixth liquid injection pump 11.3 and the seventh liquid injection pump 12.3 are opened until liquid flows out of the liquid outlet ends of the first connecting pipeline 6.2, the second connecting pipeline 7.2, the third connecting pipeline 8.2, the fourth connecting pipeline 9.2, the fifth connecting pipeline 10.2, the sixth connecting pipeline 11.2 and the seventh connecting pipeline 12.2, gas in each connecting pipeline is discharged, at the moment, the first liquid injection pump 6.3, the second liquid injection pump 7.3, the third liquid injection pump 8.3, the fourth liquid injection pump 9.3, the fifth liquid injection pump 10.3, the sixth liquid injection pump 11.3 and the seventh liquid injection pump 12.3 are closed, and the inner cavities of the first connecting pipeline 6.2, the second connecting pipeline 7.2, the third connecting pipeline 8.2, the fourth connecting pipeline 9.2, the sixth liquid injection pump 11.3 and the seventh liquid injection pump 12.3 penetrate into the reaction vessel connecting pipeline 11.2.

Step two, preparing three washing and filtering reagents: sodium hydroxide solution, purified water and acetic acid or PBS buffer solution are respectively filled into a first washing and filtering container 22, a second washing and filtering container 23 and a third washing and filtering container 24, namely, the first washing and filtering container 22 is used for containing the sodium hydroxide solution, the second washing and filtering container 23 is used for containing the purified water, and the third washing and filtering container 24 is used for containing the acetic acid or PBS buffer solution; meanwhile, the liquid inlet ends of the first washing and filtering pipeline 19, the second washing and filtering pipeline 20 and the third washing and filtering pipeline 21 correspondingly extend into the inner cavities of the first washing and filtering container 22, the second washing and filtering container 23 and the third washing and filtering container 24 one by one and are close to the bottom.

And thirdly, carrying out protein marking, adding the protein raw material solution into the reaction container 2, and controlling the constant-temperature magnetic stirrer 1 to set the corresponding required temperature for stirring and precooling.

Fourthly, starting a first liquid injection pump 6.3, filling a certain amount of sodium periodate solution in a first liquid storage container 6.2 into the reaction container 2, then automatically starting a second liquid injection pump 7.3 after timing for about 1 to 2 hours, filling a certain amount of glycol solution into the reaction container 2, and then automatically recording the weight by an electronic scale 5; then, according to the needs of protein labeling, the third liquid injection pump 8.3, the fourth liquid injection pump 9.3, the fifth liquid injection pump 10.3, the sixth liquid injection pump 11.3 or the seventh liquid injection pump 12.3 are started, so that the addition of an acetic acid solution, a hydroformylation enzyme solution, a 0.2mol/L bicarbonate buffer solution, naBH4 (sodium borohydride) or a 0.067mol/L PBS buffer solution is completed.

And fifthly, starting the electric lifting device 31, adjusting the hard straight pipe 30 to move upwards or downwards until the liquid inlet end of the hard branch pipe 30 extends into the liquid level in the reaction container 2, monitoring the liquid information in the reaction container 2 by the liquid level sensor 32 and transmitting the liquid information to the main controller, and controlling the electric lifting device 31 to stop moving by the main controller.

And sixthly, after the constant-temperature magnetic stirrer 1 works for 0.5 hour, the main controller controls the circulating filter part to work and starts a washing and filtering link. Firstly, the main controller controls the first three-position three-way valve 16 to be in a left position function (namely, the liquid inlet pipeline 14.1 is kept to be communicated with the hollow fiber column 13) and starts the circulating pump 15, at the moment, the liquid in the reaction vessel 2 is sucked into the liquid inlet pipeline 14.1 through the liquid inlet end of the hard straight pipe 30, and then enters the inner cavity through the lower port of the hollow fiber column 13 after passing through the first three-position three-way valve 16. Wherein, a part of liquid flows into the liquid outlet pipeline 14.2 from the upper port after passing through the inner cavity of the hollow fiber column 13, then sequentially passes through the third three-position three-way valve 27 and the bubble detection device 29, and then flows back to the reaction vessel 2 again; the other part of the liquid (mainly water and small molecule substances) passes through the hollow fiber membranes in the hollow fiber column 13 and flows into a collecting container 26 from a liquid collecting pipeline 25 communicated with the side wall port (i.e. the outer cavity liquid outlet of the hollow fiber column 13).

When washing and filtering, the main controller controls the third liquid injection pump 8.3 to work, and quantitatively sends the acetic acid solution contained in the third liquid storage container 8.1 (the addition amount of the acetic acid solution is determined according to the volume of the solution to be washed and filtered) into the reaction container 2.

After the circulating pump 15 runs for a certain time, the liquid level in the reaction vessel 2 will drop, when the liquid level drops below the liquid inlet end of the hard straight pipe 30, because of the continuous running of the circulating pump 15, gas will be sucked into the hard straight pipe 30 and the liquid inlet pipeline 14.1, the gas will enter the liquid outlet pipeline 14.2 through the hollow fiber column 13, in the process of flowing through the liquid outlet pipeline 14.2, the gas will pass through the bubble detection device 29, when the bubble detection device 29 detects that continuous bubbles appear for 1 minute, it is determined that the washing and filtering are completed, and the main controller can control the circulating pump 15 to stop working. Of course, the weight of the reaction vessel 2 before washing and filtering and the weight of the reaction vessel 2 after continuous bubbles appear for 1 minute can be automatically recorded by the electronic scale 5 and transmitted to the main controller, the washing and filtering volume is calculated by the main controller, and when the washing and filtering volume reaches the set volume, the main controller can control the circulating pump 15 to stop working.

And seventhly, after the hollow fiber column 13 is used up, the washing with a sodium hydroxide solution is needed, the sodium hydroxide solution can break the macromolecular protein and change the macromolecular protein into small protein fragments, so that the protein can be washed out, and the protein attached to the hollow fiber column 13 can be washed out. Firstly, the circulating pump 15 is in a standby state, the electric lifting device 31 works to lift the hard straight pipe 30 upwards to enable the liquid inlet end of the hard straight pipe to move to the position above the liquid level in the reaction vessel 2, and meanwhile, the third three-position three-way valve 27 is controlled to be in a right position (namely, the upper port of the hollow fiber column 13 is kept communicated with the liquid discharge pipeline 14.2), at the moment, the circulating pump 15 works to completely discharge the liquid remained in the hollow fiber column 13. Then the circulation pump 15 is closed, and the first three-position three-way valve 16 is adjusted to change from the left position function to the right position function, and the second three-position three-way valve 17 and the three-position five-way valve 18 are both in the left position function (i.e. the lower port of the hollow fiber column 13 is kept to be communicated with the first filter washing container 22). The circulation pump 15 is then turned on to pump the sodium hydroxide solution in the first leaching vessel 22 to wash the hollow fiber column 13, and the washed waste liquid flows into the collection vessel 26 to be collected.

And step eight, after the sodium hydroxide solution finishes washing the hollow fiber column 13, purified water washing is carried out to wash off residual protein fragments and sodium hydroxide on the hollow fiber column 13. Firstly, the circulating pump 15 stops working, and the three-position five-way valve 18 is adjusted from a left-position function to a middle-position function (namely, the second filter washing container 23 is kept communicated with the lower port of the hollow fiber column 13); the circulation pump 15 is then activated to pump purified water from the second washing and filtering vessel 23 to wash the hollow fibre column 13, and the washed waste liquid is also passed into the collection vessel 26 for collection.

And a ninth step, after the purified water washing is finished, sometimes acetic acid or PBS buffer solution washing is carried out to further wash off a small amount of protein fragments and sodium hydroxide remained on the hollow fiber column 13 and help the hollow fiber column 13 to recover the performance so as to keep the hollow fiber column 13. Firstly, the circulation pump 15 stops working, and the three-position five-way valve 18 is adjusted to the right position from the middle position function (namely, the third washing and filtering container 24 is kept communicated with the lower port of the hollow fiber column 13); the circulation pump 15 is then turned on to pump the acetic acid or PBS buffer in the third washing and filtering container 24 to wash the hollow fiber column 13, and the washed waste liquid also flows into the collecting container 16 to be collected.

After the hollow fiber column 13 is washed with acetic acid or PBS buffer solution, the cleaning of the circulating filter part after use is completed, and the reaction container 2 and each sample adding part can be directly disassembled for cleaning.

Claims (10)

1. An automated protein labeling apparatus, comprising: comprises that

The stirring reaction part consists of a constant-temperature magnetic stirrer, a reaction container arranged on the constant-temperature magnetic stirrer and a sealing cover which is matched and buckled on an opening of the reaction container;

the sample adding parts are a plurality of groups with the same structure, and each group of the sample adding parts is provided with a liquid storage container for storing a sample, a connecting pipeline for communicating the liquid storage container with the reaction container, and a liquid injection pump for conveying sample liquid into the reaction container; and

the circulating filter part is provided with a hollow fiber column for washing and filtering the sample liquid in the reaction vessel, the hollow fiber column is circularly communicated with the reaction vessel through a circulating pipeline, and the circulating pipeline is provided with a circulating pump and a washing and filtering unit.

2. The automated protein labeling apparatus of claim 1, wherein: the circulating pipeline comprises a liquid inlet pipeline and a liquid outlet pipeline, wherein the liquid inlet pipeline is communicated with the lower port of the hollow fiber column, the liquid outlet pipeline is communicated with the upper port of the hollow fiber column, and the liquid inlet end of the liquid inlet pipeline and the liquid outlet end of the liquid outlet pipeline are communicated with the inner cavity of the reaction container.

3. The automated protein labeling apparatus of claim 2, wherein: the washing and filtering unit is communicated with the liquid inlet pipeline close to the liquid outlet end, and the circulating pump is positioned on the liquid inlet pipeline between the washing and filtering unit and the hollow fiber column.

4. The automated protein labeling apparatus of claim 3, wherein: the washing and filtering unit comprises

The first three-position three-way valve is arranged on the liquid inlet pipeline;

the second three-position three-way valve is connected with the first three-position three-way valve;

the three-position five-way valve is connected with the second three-position three-way valve;

the first filter washing container is communicated with the three-position five-way valve through a first filter washing pipeline;

the second filter washing container is communicated with the three-position five-way valve through a second filter washing pipeline; and

and the third filter washing container is communicated with the three-position five-way valve through a third filter washing pipeline.

5. The automated protein labeling apparatus of claim 2, wherein: the side wall opening of the hollow fiber column is communicated with a liquid collecting pipeline, and the liquid outlet end of the liquid collecting pipeline is communicated with a collecting container.

6. The automated protein labeling apparatus of claim 5, wherein: the liquid outlet pipeline is connected with a liquid outlet pipeline through a third three-position three-way valve, the liquid outlet end of the liquid outlet pipeline is communicated with the collecting container, and a bubble detection device is arranged on the liquid outlet pipeline between the third three-position three-way valve and the reaction container.

7. The automated protein labeling apparatus of claim 2, wherein: the liquid inlet end of the liquid inlet pipeline vertically penetrates through the sealing cover to be inserted into a hard straight pipe in the reaction vessel, an electric lifting device for driving the hard straight pipe to vertically move up and down is arranged on the hard straight pipe outside the sealing cover, and a liquid level sensor which is flushed with the lower port of the hard straight pipe is arranged in the reaction vessel.

8. The automated protein labeling apparatus of claim 1, wherein: the sample adding parts are seven groups, and the liquid outlet end of the connecting pipeline of each group of sample adding parts penetrates through the sealing cover and extends into the inner cavity of the reaction container.

9. The automated protein labeling apparatus of claim 1, wherein: a PH meter for detecting the PH value is arranged in the reaction container.

10. The automated protein labeling apparatus of claim 1, wherein: and an electronic scale for weighing the weight of the reaction liquid in the reaction container is arranged at the bottom of the constant-temperature magnetic stirrer.

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