JP2005273656A - Liquid supply pump - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To make a compact liquid supplying pump system capable of eliminating the need of cleaning operation carried out for avoiding pollution during sample analysis, decreasing the risk of infection from an infection source adhered to a liquid supplying flow passage to a tester, and usable in a laboratory; and to provide a liquid supply system for simply and promptly supplying liquid without mixing of foreign matters. <P>SOLUTION: A column including a film or a gel is stored in a disposable cartridge, and a part of a side wall of the cartridge is used as a compressing plate. Part of a tube connected to the column is disposed on the side wall of the cartridge, and the tube is pressed against a drive rotation rotor to from a liquid supplying pump, thereby carrying a moving phase. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a liquid feed pump that feeds a liquid such as a stock solution containing a biological component, particularly a stock solution represented by human plasma, urine and the like.

  In the fields of biochemistry, pharmacy, cell biology, and molecular biology, for example, a specific protein is separated and purified from a biological tissue extract, or a protein in a cell culture produced by a cell is purified. An operation for separating a target substance from a mixture, such as separating a gene from a cell extract, has been performed. Means used for such operation is disclosed in Non-Patent Document 1. Of these, gel chromatography is the most versatile. Gel filtration is based on differences in molecular weight, ion exchange chromatography is based on differences in charge state, and separation is based on differences in affinity for ligands. There is affinity chromatography. In recent years, a technology for producing proteins useful for cells and bacteria has been developed due to advances in molecular biology. As shown in Patent Document 1, a target protein is separated from a culture supernatant using a hollow fiber membrane. Techniques for purification are known. In these separation technologies, a column equipped with a membrane or a column filled with gel and a liquid feed pump are connected by a silicon tube or the like, and a mobile phase such as a buffer solution is conveyed by the liquid feed pump, A stock solution containing the target substance is introduced into the membrane and passed through a membrane or column to separate the target substance.

In order to obtain the maximum separation efficiency in these techniques, it is necessary to control the speed at which the mobile phase is conveyed to a minimum. If it is too high, clogging of the film and collapsing of the gel occur, so that there is an upper limit on the conveyance speed, which is the largest cause of the time taken for these operations. When the liquid supply flow path is complicated, it takes time to assemble, and when processing a plurality of different samples, it takes more time because a washing operation is added to avoid contamination between analyses. Further, in the case where the sample is a body fluid of a patient having an infection source, the risk of the experimenter increases when the infection source adheres to the liquid delivery channel.
Japanese Patent No. 3439503 Edited by the Japanese Biochemical Society, “Seminar in Experimental Chemistry for the New Chemistry (Vol. 1) Protein (1) Separation, Purification, and Properties”, Tokyo Chemical Doujin, 1990

  In recent years, the above-described technique has been actively used in research for searching for pathological substances, and is used for processing a large amount of specimens coming from clinical sites. A compact separation system that eliminates cleaning operations to avoid contamination between analyses, reduces the risk of infection from the infection source attached to the flow path to the experimenter, and can be used in the laboratory. It is a problem to be solved to create a liquid feed pump that can cope with this.

  As a result of studies to solve the above problems, a column containing a membrane or gel is housed in a disposable cartridge, and a part of the cartridge side wall (a part of the outer shell of the cartridge) is used as a pressing plate. Then, a part of the tube connected to the column was placed on the side wall of the cartridge and the tube was pressed against the driving rotary rotor, and it was found that a liquid feed pump was formed and the mobile phase could be conveyed, leading to the present invention.

According to the preferable form of this invention, following (1)-(15) can be mentioned.
“(1) A driving rotary rotor, a tube forming a liquid feeding flow path, and a pressing plate, and the tube is pressed against the pressing plate by the driving rotary rotor and rotated in the circumferential direction of the tube. A liquid feed pump for feeding an internal liquid, wherein the squeezing plate, which is a component of the liquid feed pump, is a part of an outer shell of a cartridge detachable from the liquid feed pump .
(2) The liquid feed pump according to (1), wherein the tube which is a component of the liquid feed pump is a part of the liquid feed flow path installed in the cartridge.
(3) The liquid feed pump according to (1) or (2), wherein a tube is installed on a driving rotary rotor pressing surface of the cartridge outer shell.
(4) The liquid feeding pump according to any one of (1) to (3), wherein the driving rotary rotor pressing surface of the cartridge outer shell forms a curved surface that is curved in an arc shape in a pressing direction.
(5) The liquid feed pump according to (4), wherein the tube is installed in an arcuate shape with respect to the drive rotary rotor pressing surface bent in an arc shape of the cartridge outer shell.
(6) The liquid feed pump according to any one of (1) to (5), wherein at least one column is connected to the liquid feed flow path.
(7) The liquid feeding pump according to (6), wherein a membrane is installed on the column.
(8) The liquid feed pump according to (7), wherein the membrane is a hollow fiber membrane.
(9) The material of the membrane is selected from cellulose, cellulose acetate, polycarbonate, polysulfone, polymethacrylate, polyacrylate, polyamide nylon, polyvinylidene fluoride, polyacrylonitrile, polyester, polyurethane, polystyrene, polyethylene and polypropylene (7) or (8) The liquid feeding pump as described.
(10) The liquid feed pump according to (6), wherein the column is a gel packed column.
(11) The liquid feed pump according to (6), wherein the column is a column for performing gel filtration or adsorption separation.
(12) The liquid feeding pump according to any one of (6) to (11), wherein at least one membrane column and at least one gel packed column are connected to the liquid feeding flow path.
(13) The liquid feeding pump according to any one of (1) to (12), wherein the liquid feeding channel is a part of a closed channel.
(14) The liquid feeding pump according to any one of (1) to (13), wherein all the liquid feeding flow paths are housed in a cartridge.
(15) The liquid feed pump according to any one of (1) to (14), which is provided with a mechanism for transporting the cartridge to a position where the drive rotary rotor can squeeze the tube installed in the cartridge when the cartridge is installed.

  By separating the squeeze plate and tube that make up the liquid delivery pump from the drive rotor, and using a disposable cartridge containing the separation column as the squeeze plate, it is possible to save the trouble of assembling the liquid delivery flow path and to remove the cartridge for each sample analysis. It only needs to be exchanged, and the washing operation of the liquid supply flow path, which is performed to avoid contamination between analyses, becomes unnecessary. Since the infection source adhering to the liquid flow path does not accumulate in the liquid flow path, the risk of infection to the experimenter is extremely reduced. In addition, it will be a compact pump system that can be used in laboratories.

  The cartridge referred to in the present invention is a box having a storage space. The material is not particularly limited, but a plastic material is preferable because it is easy to handle, transport, and has strength. The shape is not particularly limited, but there is enough space inside to accommodate the column and the liquid supply flow path, and the pressing surface of the squeezing plate that is squeezed by the driving rotary rotor of the liquid supply pump is arcuate in the direction to be compressed It is particularly preferable that the curved surface is curved. More preferably, all the liquid supply flow paths are housed in the cartridge. The curved surface increases the contact area with the drive rotor, and as a result, a stable flow rate can be secured. This outer shell portion corresponds to a pressing plate that is a component of the liquid feed pump. It is preferable that the tube which is another component of a liquid feeding pump is installed in this pressing surface. A liquid feed pump is formed in such a manner that the compressed surface of the cartridge and the drive rotary rotor sandwich the tube, and the liquid in the liquid feed flow path in the cartridge circulates as the drive rotary rotor rotates in the circumferential direction. Although it is essential that the tube be installed in the direction in which the tube is squeezed with respect to the squeezed surface of the cartridge outer shell, the tube may not necessarily be in contact with the squeezed surface. In order to prevent the tube from vibrating in a direction perpendicular to the squeezing direction during pressing, it is particularly preferable that the tube is installed in an arcuate shape with respect to the pressing surface that is bent in an arc shape of the outer shell of the cartridge.

  The drive rotating rotor of the present invention is preferably provided with a rotatable roller on its outer periphery. If there is a roller on the outer periphery, the tube can be squeezed more smoothly. The amount of mobile phase delivered by the tube pump can be determined by the amount of mobile phase present in the unsqueezed tube between the rollers.

  The liquid feeding flow path of the present invention refers to a path through which a specimen to be processed and a mobile phase such as water or a buffer are transported. The liquid supply flow path through which the mobile phase is supplied by the liquid supply pump of the present invention is not particularly limited, and any liquid supply flow path may be used as long as a tube that is squeezed by the drive rotating rotor is installed. As will be described later, a liquid feeding flow path including a column for separating biological components is preferably used. The end of the liquid feeding flow path may be released or may be a closed flow path forming a closed loop, but is preferably a closed flow path. By being a closed flow path, all the liquid supply flow paths can be stored in the cartridge. When a column is installed, it may be installed anywhere in the liquid feed channel. The material of the tube is not particularly limited, but it is preferably made of a flexible elastic body, and silicone resin, polyvinyl chloride, polyurethane, fluororesin, natural rubber, and synthetic rubber are preferably used, and are durable and stable. Most preferred are silicone resins and fluororesins.

  The direction in which the tube is attached to the squeezed surface of the cartridge matches the direction of the column port. Also, the same number of grooves as the column are formed on the surface of the outer shell of the cartridge that is squeezed, and the tube can be firmly fixed by filling the groove with the tube. The tubes connected to the both end ports of each column can circulate the liquid in the liquid feeding flow path through the pressing plate. In order to maintain the accuracy of pressing, the tube is preferably located closer to the support (machine side). If the accuracy goes wrong, the tube cannot be pressed down and fixed quantity feeding becomes impossible. In order to easily and accurately mount the cartridge on the main body, the cartridge is provided with a guide hole, and the guide hole is passed through the guide shaft of the pump drive device, so that the cartridge is easily mounted. Subsequently, by fixing the cartridge, the plurality of tubes are kept at an appropriate distance from the driving rotation rotor of the liquid feeding pump, and by operating the liquid feeding pump, the plurality of hollow fiber column processing liquids are sequentially fed. Is done.

  In the present invention, a liquid flow path connected to a column having a hollow fiber membrane or a flat membrane is preferably used, and a hollow fiber membrane is more preferably used because of a high throughput and a small pressure loss. . Regarding the molecular fractionation performance of the membrane, the molecular fractionation ability (cutoff value) can be appropriately selected in consideration of the molecular weight of the solute to be collected and the molecular weight of the solute to be removed. It is preferable that the drive rotor of the liquid feed pump squeezes the tube constituting the liquid feed flow path, and the treatment liquid can be circulated through the hollow fiber in the column by the liquid feed pump.

  For example, when a multi-stage configuration is used to increase the separation efficiency, a plurality of liquid feeding channels are arranged horizontally, and the filtrate outlet of the preceding hollow fiber column and the latter feeding channel are connected by a feeding pipe. Then, the treatment liquid is sequentially sent to a multistage column, and separation is repeated. In order to send the liquid without stagnation and obtain the maximum separation efficiency, it is preferable that the drive rotary rotor squeezes each tube of the multistage liquid feeding flow path. These pumps may be operated with separate drives or coaxially operated with the same drive, but are operated so as to maintain the same flow rate in order to send liquid without stagnation and obtain the maximum separation efficiency. It is preferred that When operating on the same axis, the operating speed and sequence of each part can be appropriately selected according to the separation / concentration efficiency of the membrane.

  In the present invention, the membrane used for filtration is a group consisting of cellulose, cellulose acetate, polycarbonate, polysulfone, polymethacrylate, polyacrylate, polyamide nylon, polyvinylidene fluoride, polyacrylonitrile, polyester, polyurethane, polystyrene, polyethylene and polypropylene. The target solute component can be more efficiently separated by using a filter or hollow fiber containing one or more materials selected from the above. Any of flat membrane type separation membranes (filters) such as flat filters and cartridge type filters, and hollow separation membranes (hollow fibers) such as hollow fibers can be used.

These filters or hollow fibers contain antibodies and fragments thereof, polyethyleneimine, aminomethylpyridine, polyphenol, blue dye, divalent metal ions (Zn ² +, Ni ² +, Co ² +, Cu ² +, etc.) and hydrophobicity By immobilizing one or more substances (ligands) selected from the group consisting of compounds (methyl group, benzyl group, phenyl group, chloromethyl group, octyl group, lauryl group, etc.), the solute is added to the filter or hollow fiber. Affinity can be imparted.

  The cartridge may be pressed directly against the drive rotating rotor, but from the point of safety to the operator, the cartridge moves when the cartridge is installed, and the cartridge is transported to a position where the rotor can squeeze the tube installed in the cartridge It is preferable to provide.

  Hereinafter, one embodiment of the method for separating protein components in an aqueous solution and the protein fractionation apparatus of the present invention will be described with reference to the drawings, but the present invention is not limited thereto.

(Example)
1, 2 and 3 are diagrams of an example of a fractionation apparatus provided with the liquid feeding pump of the present invention. FIG. 1 shows that the separation part is constituted by a three-stage liquid flow path. A three-way joint 2a and a joint 2c were connected to the rubber button 2b. Using the flexible tube 3, the joint 2 c and the lower nozzle 6 a of the hollow fiber membrane column 5 a were connected so as to run over the curved surface of the multichannel pressing plate 8. Further, the three-way joint 2a was connected to the back 12 with a tube. The hollow fiber membrane columns 5a, 5b, 5c constituting the separation part, and the hollow fiber membrane column 5d constituting the concentrating part and the multi-channel type pressing plate 8 are integrated into each hollow fiber membrane column 5a, 5b, 5c, 5d. A flexible tube is connected to the upper nozzles 4a, 4b, 4c, and 4d provided at the upper end of each, and each tube is installed so as to run over the curved surface of the multi-channel type compression plate 8, and the lower nozzles 6a, 6b, 6c, Each connected to 6d.

  Next, the lower body nozzle 7a of the hollow fiber membrane column 5a and the lower nozzle 6b of the hollow fiber membrane column 5b, and the lower body nozzle 7b of the hollow fiber membrane column 5b and the lower nozzle 6c of the hollow fiber membrane column 5c are hollow. The trunk lower nozzle 7c of the yarn membrane column 5c and the lower nozzle 6d of the hollow fiber membrane column 5d were connected by piping. The trunk lower nozzle 7d of the hollow fiber membrane column 5d and the three-way joint 2a were connected by piping. Furthermore, the lower nozzle 6d of the hollow fiber membrane column 5d and the recovery container cap 11 were connected, and the upper nozzle 4d of the hollow fiber membrane column 5d and the recovery container cap 11 were connected. The recovery container cap 11 of the recovery container 10 was sealed. All the hollow fiber membrane columns, nozzles, tubes, joints, backs with tubes, collection containers and collection container caps described above formed closed channels. The closed flow path was filled with an aqueous buffer which is a mobile phase.

  As shown in FIG. 2, the liquid feeding flow path is housed in a housing box 13 to form a cartridge C. The device 14 is equipped with a multi-channel drive rotary rotor 9, and the cartridge and the guide hole of the multi-channel pressing plate 8 penetrate the guide shafts 8a and 8b and are fixed by being pushed into the guide shafts 8a and 8b. The fixed cartridge C moved in parallel to the drive rotation rotor 9 side, and a liquid feed pump was formed by the drive rotation rotor 9, the compression plate 8, and seven tubes installed on the curved surface of the compression plate 8. Eight rotating rollers that can freely rotate were installed on the outer periphery of the drive rotating rotor 9.

  As shown in FIG. 3, a syringe pump S was attached to the upper part of the multi-channel drive rotating rotor 9. A motor was attached to each of the multi-channel drive rotary rotors 9.

  The liquid flow is indicated by arrows. After inserting the needle of the syringe 1 enclosing a stock solution such as serum into the rubber button 2b of the supply unit, the specimen was introduced at a speed controlled by a syringe pump. The administered stock solution was fed to the hollow fiber membrane column 5a of the separation unit while being mixed with the mobile phase and being transported by a driving rotary rotor 9a driven by a motor. The filtrate produced while circulating through the hollow fiber membrane column 5a by the drive rotary rotor 9b driven by the motor exited from the lower fuselage nozzle 7a and was conveyed to the subsequent hollow fiber membrane column 5b by the drive rotary rotor 9b. The filtrate of the hollow fiber membrane column 5b was further transported to the subsequent hollow fiber membrane column 5c. Thus, the solute of the undiluted solution was fractionated by the hollow fiber membrane columns 5a, 5b and 5c forming the separation part. The filtrate of the hollow fiber membrane column 5c was transported to the hollow fiber membrane column 5d of the concentrating part. The filtrate produced while circulating through the hollow fiber membrane column 5d exited from the fuselage lower nozzle 7a and was returned to the supply section via the joint 2a. The filtrate of the hollow fiber membrane column 5c was transported to the hollow fiber membrane column 5d of the concentrating part. The liquid circulation and liquid feeding in the separation unit and the concentration unit were performed by a driving rotary rotor 9b. After the designated time has elapsed, the driving rotary rotors 9a and 9b are stopped, and the driving rotary rotor 9c driven by the motor is started. As a result, the air in the recovery container 10 pushed out the concentrated liquid in the liquid flow path in the concentration section, and the concentrated liquid was recovered in the recovery container 10 through the lower nozzle 6d. Note that Ga and Gb in FIG. 3 are the same as 8a and 8b in FIGS. 1 and 2, respectively.

It is explanatory drawing which shows an example of the liquid feeding pump system which consists of the liquid feeding flow path which consists of a supply part, a separation part, a supply part, and a collection | recovery part, a pressing board, and a drive rotation rotor. It is explanatory drawing which shows a mode that the closed flow path of FIG. 1 was accommodated in the storage box, and it was set as the cartridge. It is explanatory drawing which shows an example of the liquid feeding pump apparatus of this invention. It is detailed explanatory drawing which shows a mode that the closed flow path of FIG. 1 was accommodated in the storage box, and it was set as the cartridge. It is detailed explanatory drawing which shows an example of the liquid feeding pump apparatus of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Syringe 2a 3 way joint 2b Rubber button 2c Joint 3 Tube 4a Upper nozzle 4b Upper nozzle 4c Upper nozzle 4d Upper nozzle 5a Hollow fiber membrane column 5b Hollow fiber membrane column 5c Hollow fiber membrane column 5d Hollow fiber membrane column 6a Lower nozzle 6b Below Nozzle 6c Lower nozzle 6d Lower nozzle 7a Body lower nozzle 7b Body lower nozzle 7c Body lower nozzle 7d Body lower nozzle 8a Guide shaft 8b Guide shaft 9 Multi-channel drive rotary rotor 9a Drive rotary rotor 9b Drive rotary rotor 9c Drive rotary rotor 10 Recovery Container 11 Collection container cap 12 Back with tube 13 Storage box 14 Device 15 Rotating roller around drive rotor

Claims (15)

It consists of a drive rotating rotor, a tube forming a liquid feeding flow path, and a pressing plate, and the tube is pressed against the pressing plate by the driving rotating rotor, and the liquid in the tube is removed by circumferential rotation of the driving rotating rotor. A liquid feed pump for feeding liquid, wherein a pressing plate, which is a component of the liquid feed pump, is a part of an outer shell of a cartridge that can be attached to and detached from the liquid feed pump. The liquid feed pump according to claim 1, wherein the tube which is a component of the liquid feed pump is a part of a liquid feed flow path installed in the cartridge. The liquid feeding pump according to claim 1 or 2, wherein a tube is installed on a driving rotary rotor pressing surface of the cartridge outer shell. The liquid feed pump according to any one of claims 1 to 3, wherein a drive rotary rotor pressing surface of the cartridge outer shell forms a curved surface that is curved in an arc shape in a pressing direction. The liquid feeding pump according to claim 4, wherein the tube is installed in an arcuate shape with respect to the driving rotary rotor pressing surface bent in an arc shape of the cartridge outer shell. The liquid feeding pump according to claim 1, wherein at least one column is connected to the liquid feeding flow path. The liquid feed pump according to claim 6, wherein a membrane is installed on the column. The liquid feeding pump according to claim 7, wherein the membrane is a hollow fiber membrane. 9. The material according to claim 7, wherein the material of the membrane is selected from cellulose, cellulose acetate, polycarbonate, polysulfone, polymethacrylate, polyacrylate, polyamide nylon, polyvinylidene fluoride, polyacrylonitrile, polyester, polyurethane, polystyrene, polyethylene and polypropylene. Feed pump. The liquid feed pump according to claim 6, wherein the column is a gel packed column. The liquid feed pump according to claim 6, wherein the column is a column for performing gel filtration or adsorption separation. The liquid delivery pump according to any one of claims 6 to 11, wherein at least one membrane column and at least one gel packed column are connected to the liquid delivery flow path. The liquid feeding pump according to claim 1, wherein the liquid feeding channel is a part of a closed channel. The liquid feeding pump according to claim 1, wherein all the liquid feeding passages are housed in a cartridge. The liquid feed pump according to any one of claims 1 to 14, further comprising a mechanism for transporting the cartridge to a position where the drive rotary rotor can squeeze the tube installed in the cartridge when the cartridge is installed.

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