JP2016156843A - Biomolecule analysis apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a vertical-type electrophoresis-transfer apparatus of an ejection transfer system, having an appropriate transfer film conveyance mechanism.SOLUTION: A biomolecule analysis apparatus (100) includes: an arm part (20-23, 66) holding a transfer film (1) arranged at a position facing a first opening (50a); and a drive part (62-65) arranged under an anode buffer tank 30 and configured to drive the arm part substantially in horizontal direction. The arm part is connected to the drive part (62-65) and configured to be driven such that a sample separated from the first opening (50a) is transferred onto the transfer film (1) at a timing of ejection.SELECTED DRAWING: Figure 1

Description

  The present invention relates to an apparatus for analyzing biomolecules, and more particularly to an apparatus for transferring biomolecules separated by electrophoresis to a transfer film.

  In Patent Documents 1 and 2, a device for electrophoresis of biomolecules such as DNA and protein in a gel and transferring it directly from the gel end face to a transfer film (hereinafter referred to as “discharge transfer electrophoresis”). Sometimes referred to as "transfer device").

  The devices described in Patent Documents 1 and 2 are a gel arranged in a horizontal direction, a gel frame for storing the gel, a cathode buffer tank and an anode buffer tank respectively connected to both ends of the gel, and opposed to one end of the gel. And a rectangular transfer film frame that fixes the transfer film, and a vertical transport mechanism that transports the transfer film frame in the vertical direction. The horizontal apparatus transfers DNA or the like separated in the gel to the transfer film by performing the electrophoresis in the gel and pulling up the transfer film by the vertical transport mechanism.

US Pat. No. 5,234,559 (registration date: August 10, 1993) US Pat. No. 5,916,429 (registration date: June 29, 1999)

  However, in the conventional horizontal apparatus, the gel frame is provided in the horizontal direction and the transfer film is pulled up in the vertical direction (this configuration is also referred to as “horizontal type” in the present specification). In the horizontal apparatus, the upper surface of the gel frame is in contact with the outside air, and the lower surface of the gel frame is in contact with the structure of the apparatus. Therefore, the gel frame is not sufficiently cooled, and the gel may generate heat during electrophoresis. If the gel generates heat, the electrophoretic pattern may be distorted, the resolution may be reduced, and the separated DNA may not be transferred to the transfer film. The transfer film is generally a thin plastic film and has elasticity. Therefore, the film fixed to the rectangular transfer film frame is in a state where the center is more easily extended than the peripheral part, and the tension is increased as it approaches the edge. For this reason, the transfer pattern may be distorted near the center.

  Therefore, the present inventors examined a vertical apparatus having a unique configuration based on a unique idea. In the vertical apparatus, a separation unit that stores a separation medium such as a gel stands up in a substantially vertical direction (this configuration is also referred to as a “vertical type” in the present specification). In the vertical apparatus, the upper part of the separation part is in contact with the cathode buffer and the lower part of the separation part is in contact with the anode buffer, so that the separation part can be sufficiently cooled by water cooling. The transfer film can be transported without being fixed to the rectangular transfer film frame.

However, in the vertical apparatus, it is necessary to move the transfer film in a substantially horizontal direction in the anode buffer tank. Here, when the transfer film transport mechanism is arranged upstream of the transfer film as in the conventional horizontal apparatus, the buffer solution scattered from the anode buffer tank reduces the durability of the transfer film transport mechanism. And there is a risk that the transfer film transport mechanism hinders various operations on the vertical apparatus.

  The present invention has been made in view of the above-described problems, and has as its main object to provide a vertical discharge transfer type electrophoresis-transfer apparatus equipped with a suitable transfer film transport mechanism.

  In order to solve the above problems, a biomolecule analyzer according to an aspect of the present invention stores a first buffer tank, a second buffer tank disposed above the first buffer tank, and a separation medium. The first opening that opens in the first buffer solution tank and the second opening that opens in the second buffer solution tank, and is arranged at a position facing the first opening, a separation portion that stands in a substantially vertical direction. An arm portion for holding the transfer film, and a drive portion that is provided under the first buffer solution tank and drives the arm portion in a substantially horizontal direction, the arm portion being connected to the drive portion, It is driven so that the sample separated from one opening is transferred to the transfer film at the timing when it is discharged.

  In addition, in order to solve the above problems, a biomolecule analyzer according to an aspect of the present invention stores a first buffer solution tank, a second buffer solution tank disposed above the first buffer solution tank, and a separation medium. A separation portion that has a first opening that opens into the first buffer solution tank and a second opening that opens into the second buffer solution tank, and stands in a substantially vertical direction, at a position facing the first opening. An arm portion for holding the transfer film disposed, and a drive portion that is provided under the first buffer solution tank and drives the arm portion in a substantially horizontal direction. The arm portion is provided on a side wall of the first buffer solution tank. It passes through the outside, wraps around the upper end of the side wall, and is connected to the inside of the side wall.

  According to said structure, by setting it as the structure which a isolation | separation part stands substantially perpendicularly, a isolation | separation part can be immersed in the buffer solution in a 1st or 2nd buffer solution tank, and a separation medium can be water-cooled. However, when the biomolecule analyzer is configured in this way, it is necessary to move the transfer film in the first buffer tank.

  Here, according to said structure, a drive part is provided under a 1st buffer solution tank, and the shape of an arm part passes along the outer side of the side wall of a 1st buffer solution tank, and wraps around the upper end of the said side wall. By adopting a shape that is connected to the inside of the side wall, the transfer film can be successfully moved in the first buffer solution tank while avoiding deterioration of the durability of the drive unit due to the buffer solution and obstruction of various operations by the drive unit. Can be made. Accordingly, it is possible to provide a vertical discharge transfer type electrophoresis-transfer apparatus including a suitable transfer film transport mechanism.

  In the biomolecule analyzer according to one aspect of the present invention, the transfer film has a first end portion in the moving direction forward by driving the arm portion and a second end portion in the moving direction rear, and the arm portion is A first fixing portion that fixes the first end portion, a second fixing portion that fixes the second end portion, and an elastic body that biases the first fixing portion and the second fixing portion in a direction facing each other. It may be.

  If the transfer film is loose, when the transfer film is moved, the gap between the transfer film and the first opening may become large and the transfer result may be blurred.

  According to the above configuration, the elastic body biases the first and second fixing portions that fix both ends in the moving direction of the transfer film in the directions opposite to each other, so that a constant tension is applied to the transfer film. Can be granted. Thereby, the transfer film can be stretched and a good transfer result can be obtained.

  In the biomolecule analyzer according to one aspect of the present invention, a support member that supports the transfer film from the side opposite to the separation part in the transfer film is provided at the bottom of the first buffer tank. The transfer film may be bent by the separation portion so that the opposite side to the separation portion is convex.

  According to the above configuration, the transfer film is supported by the support member, and the separation unit presses the transfer film and bends the projection downward (toward the side opposite to the separation unit). As a result, tension is applied to the transfer film, and the transfer film can be brought into close contact with the first opening. In particular, since the transfer film is maintained in a tensioned state by the elastic body, the transfer film can be suitably pressed against the first opening. Thereby, the transfer from the separation medium to the transfer film can be more suitably performed.

  In the biomolecule analyzer according to one aspect of the present invention, the support member may be formed at a position sandwiching a pair of positions facing the first opening at the bottom.

  According to the above configuration, the transfer film is supported by the support members disposed on both sides of the separation unit, and the separation unit holds the transfer film and bends it downward so as to be convex (opposite to the separation unit). . As a result, a uniform tension is applied to the transfer film, and the transfer film can be uniformly adhered to the first opening. Thereby, the transfer from the separation medium to the transfer film can be more suitably performed.

  In the biomolecule analyzer according to one aspect of the present invention, the transfer film has a first end portion in the moving direction forward by driving the arm portion and a second end portion in the moving direction rear, and the arm portion is A first fixing portion that fixes the first end portion, a second fixing portion that fixes the second end portion, and a connecting portion that connects the first fixing portion and the second fixing portion with a predetermined distance therebetween. Good.

  According to said structure, by fixing the 1st and 2nd edge part of a transfer film by the 1st and 2nd fixing | fixed part connected with predetermined distance separation, respectively. The transfer film can be stretched without loosening along its moving direction. Thereby, it can suppress that a transcription | transfer result shakes by the looseness of a transfer film, and can improve a measurement sensitivity.

  In the biomolecule analyzer according to one aspect of the present invention, the connecting portion may be disposed at a position sandwiching the transfer film from the side in the movement direction.

  According to said structure, it can avoid that a connection part overlaps the surface (surface facing 1st opening) and back surface (surface on the opposite side to 1st opening) of a transfer film. Accordingly, it is possible to prevent the transfer from the separation medium to the transfer film and the contact of other members with the back surface of the transfer film from being hindered by the connecting portion.

  In the biomolecule analyzer according to one aspect of the present invention, a support member that supports the transfer film from the side opposite to the separation part in the transfer film is provided at the bottom of the first buffer tank. The transfer film may be bent by the separation portion so that the opposite side to the separation portion is convex.

  According to the above configuration, the transfer film is supported by the support member, and the separation unit presses the transfer film and bends the projection downward (toward the side opposite to the separation unit). As a result, tension is applied to the transfer film, and the transfer film can be brought into close contact with the first opening. Thereby, the transfer from the separation medium to the transfer film can be more suitably performed.

  In the biomolecule analyzer according to one aspect of the present invention, the support member may be formed at a position sandwiching a pair of positions facing the first opening at the bottom.

  According to the above configuration, the transfer film is supported by the support members disposed on both sides of the separation unit, and the separation unit holds the transfer film and bends it downward so as to be convex (opposite to the separation unit). . As a result, a uniform tension is applied to the transfer film, and the transfer film can be uniformly adhered to the first opening. Thereby, the transfer from the separation medium to the transfer film can be more suitably performed.

  In the biomolecule analyzer according to an aspect of the present invention, the inclination angle from the position in contact with the support member to the position in contact with the first opening in the transfer film is 1 ° or more and 60 ° or less downward with respect to the horizontal plane. There may be.

  According to said structure, the tension | tensile_strength concerning a transfer film can be adjusted appropriately, and the transfer from a separation medium to a transfer film can be performed more suitably.

  In the biomolecule analyzer according to one aspect of the present invention, the portion of the arm portion that goes around the upper end of the side wall is detachable from the drive unit, and the first buffer solution tank is detachable from the biomolecule analyzer. It may be possible.

  According to said structure, since a 1st buffer solution tank can be removed, a 1st buffer solution tank can be wash | cleaned easily, without making a drive liquid adhere to a drive part. Further, when removing the first buffer solution tank, it is possible to separate the portion connected to the inner side of the side wall of the arm part from the upper end of the side wall of the first buffer solution tank from the drive unit. One buffer tank can be easily removed.

  In the biomolecule analyzer according to one aspect of the present invention, the arm unit is connected to the drive unit, and a first portion extending outside the side wall to a position aligned with an upper end of the side wall; You may have the 2nd site | part which fits into one site | part and extends inside the said side wall across the upper end of the said side wall.

  According to said structure, a 2nd site | part can be easily attached or detached with respect to a drive part. The first part is disposed outside the side wall of the first buffer tank, and does not obstruct various operations such as removal of the first buffer tank and setting of the electrodes. Therefore, various operations can be successfully performed by appropriately removing the second part.

  In the biomolecule analyzer according to one aspect of the present invention, the first buffer solution tank, the second buffer solution tank, and the separation unit may be transparent.

  According to the above configuration, the state of the separation medium and the transfer film can be observed during the operation of the apparatus. Thereby, for example, the movement of the visible marker can be confirmed with the naked eye.

  In the biomolecule analyzer according to one aspect of the present invention, the first electrode is disposed in the first buffer solution tank, the second electrode is disposed in the second buffer solution tank, and the transfer film has the first opening and You may arrange | position so that it may be inserted in a 1st electrode.

  According to the above configuration, a voltage can be applied to the separation medium between the first opening opened in the first buffer solution tank and the second opening opened in the second buffer solution tank. Molecular electrophoresis can be performed successfully. In addition, since the transfer film is inserted between the first opening and the first electrode, it is possible to successfully transfer the separated biomolecule from the first opening to the transfer film.

  In the biomolecule analyzer according to one aspect of the present invention, the separation unit is detachably attached to the second buffer solution tank, and the second buffer solution tank is attached to and detached from the first buffer solution tank. It may be attached as possible.

  According to the above configuration, since the separation unit and the second buffer solution tank can be removed, the separation unit and the second buffer solution tank can be easily washed without attaching a cleaning solution or the like to the drive unit.

  According to the present invention, it is possible to provide a vertical discharge transfer type electrophoresis-transfer apparatus having a suitable transfer film transport mechanism.

1 is a perspective view showing a schematic configuration of a biomolecule analyzer according to an embodiment of the present invention. It is sectional drawing which shows schematic structure of the biomolecule analyzer which concerns on one Embodiment of this invention. It is a perspective view which shows schematic structure of the adjuster in one Embodiment of this invention. It is sectional drawing for demonstrating the electrophoresis and transcription | transfer of the sample in one Embodiment of this invention. It is a perspective view which shows the mode of removal of each member of the biomolecule analyzer which concerns on one Embodiment of this invention. It is a perspective view which shows the mode of removal of each member of the biomolecule analyzer which concerns on one Embodiment of this invention. It is a perspective view which shows the mode of removal of each member of the biomolecule analyzer which concerns on one Embodiment of this invention. It is a perspective view which shows the mode of removal of each member of the biomolecule analyzer which concerns on one Embodiment of this invention. 1 is a perspective view showing a schematic configuration of a biomolecule analyzer according to an embodiment of the present invention. It is sectional drawing which shows schematic structure of the adjuster in one Embodiment of this invention.

Embodiment 1
An embodiment of the present invention will be described below with reference to the drawings.

  First, a schematic configuration of the biomolecule analyzer 100 according to the present embodiment will be described with reference to FIGS. 1 and 2. FIG. 1 is a perspective view schematically showing the configuration of the biomolecule analyzer 100. FIG. 2 is a side sectional view schematically showing the configuration of the biomolecule analyzer 100.

  As shown in FIGS. 1 and 2, the biomolecule analyzer 100 is a vertical discharge transfer type electrophoretic transfer device, which includes a clamp (arm unit, first fixing unit) 20, a clamp (arm unit, first unit). Two fixed parts) 21, a clamp frame (arm part, connecting part) 22, a carrier (arm part, a part that goes around the upper end of the side wall, a second part) 23, an anode buffer tank (first buffer liquid tank) 30, a table 31, Cathode buffer tank (second buffer solution tank) 40, separation part 50, motor (drive part) 62, ball screw (drive part) 63, guide shaft (drive part) 64, shaft holder (drive part) 65, guide pole (arm) Part, first part) 66, and control unit 68. Further, although not shown for the sake of explanation, for the sake of safety, a lid that covers the whole during operation is further provided.

  Here, the separation unit 50 accommodates a separation gel (separation medium) 52, and has a first opening 50 a that opens into the anode buffer tank 30 and a second opening 50 b that opens into the cathode buffer tank 40. Yes. Further, the transfer film 1 is disposed in the anode buffer tank 30 so as to face the first opening 50a. An anode (first electrode) 32 is disposed in the anode buffer tank 30, and a cathode (second electrode) 41 is disposed in the cathode buffer tank 40.

  For this reason, in the biomolecule analyzer 100, the cathode buffer tank 40 and the anode buffer tank 30 are filled with a buffer solution, so that the cathode 41 in the cathode buffer tank 40 and the anode 32 in the anode buffer tank 30 are two tanks. Are electrically connected via the buffer solution, the separation gel 52, and the transfer film 1. That is, the biomolecule analyzer 100 applies a voltage between the cathode 41 and the anode 32 to separate the sample introduced from the second opening 50b by the separation gel 52, and separates each separated component into the first. This is an apparatus for discharging from the opening 50a and adsorbing it to the transfer film 1.

  Below, each main member is demonstrated in detail with reference to FIG.1 and FIG.2.

(Anode and cathode)
The anode 32 is disposed in the anode buffer tank 30, and the cathode 41 is disposed in the cathode buffer tank 40. The anode 32 and the cathode 41 are formed from a conductive material such as metal. As a material for forming the anode 32 and the cathode 41, for example, platinum is preferable from the viewpoint of suppressing ionization of the electrode.

  Regarding the electrode arrangement, the anode 32 is arranged in the anode buffer tank 30 and the cathode 41 is not particularly limited as long as it is arranged in the cathode buffer tank 40. For example, the cathode 41, the first opening 50a and the anode 32 may be arranged on a substantially straight line. If the transfer film 1 is arranged in such an arrangement as shown in FIG. 1, the lines of electric force passing through the first opening 50a are substantially perpendicular to the transfer film 1, thereby improving the accuracy of sample adsorption. obtain.

  The anode 32 is preferably arranged away from the transfer film 1. Thereby, it is possible to suppress the bubbles generated from the anode 32 from adversely affecting the adsorption of the separation component to the transfer film 1.

  For example, the anode 32 and the cathode 41 may be used by being connected to the control unit 68 or may be used by being connected to an external power supply (DC power supply device). When used by connecting to an external power supply, after setting the time, current and voltage to the power supply, the biomolecule analyzer 100 is started by operating the control unit 68 simultaneously with the start of the operation of the power supply. Just do it.

(Anode buffer tank and cathode buffer tank)
The anode buffer tank 30 and the cathode buffer tank 40 are insulating containers for retaining a buffer solution (buffer). The cathode buffer tank 40 is provided above the anode buffer tank 30. In this embodiment, the anode buffer tank 30 is fixed on the table 31, and the cathode buffer tank 40 is fixed to the anode buffer tank 30, but the present invention is not limited to this configuration.

  The buffer solution put into the anode buffer tank 30 and the cathode buffer tank 40 can be any buffer solution having conductivity, and in particular, a buffer solution having a weakly acidic to weakly basic buffer region can be suitably used. Examples of such buffers include Tris / glycine buffer, acetate buffer, sodium carbonate buffer, CAPS buffer, Tris / boric acid / EDTA buffer, Tris / acetic acid / EDTA buffer, MOPS, Buffers such as phosphate buffer and Tris / Tricine buffer can be used.

  As will be described in detail later, the transfer film 1 is supported on the bottom of the anode buffer tank 30 from the back surface (the surface opposite to the separation portion 50) of the transfer film 1 in the movement path of the transfer film 1. Guides (support members) 33 and 34 are provided.

(Separation part)
The separation unit 50 houses a separation gel (separation medium) 52 therein. In the present embodiment, the separation unit 50 stands up in a substantially vertical direction, the lower part thereof is arranged in the anode buffer tank 30, and the upper part thereof is arranged so that one side thereof is in contact with the cathode buffer tank 40. Thereby, the separation gel 52 is water-cooled by at least one of the buffer solution in the anode buffer tank 30 and the buffer solution in the cathode buffer tank 40 and can be sufficiently cooled.

  The separation unit 50 has a first opening 50 a that opens into the anode buffer tank 30 and a second opening 50 b that opens into the cathode buffer tank 40. Thereby, the separation gel 52 faces the anode buffer tank 30 through the first opening 50a, and faces the cathode buffer tank 40 through the second opening 50b. In the present embodiment, the separation unit 50 is fixed to the cathode buffer tank 40 by a lock 42 provided in the cathode buffer tank 40, but the present invention is not limited to this configuration.

  The separation part 50 may be composed of two insulating plates 51 and 53 formed of an insulator such as glass or acrylic. In one embodiment, the separation part 50 exposes the separation gel 52 by lacking a part of the insulating plate 53 in the second opening 50 b, thereby easily introducing the sample into the separation gel 52. be able to.

  The separation gel 52 is a gel for separating the sample components introduced from the second opening 50b according to the molecular weight. The separation gel 52 can be filled in the separation unit 50 before or after the separation unit 50 is attached to the biomolecule analyzer 100. A commercially available page chip filled with the separation gel 52 may be used as the separation unit 50. Examples of the separation gel 52 include acrylamide gel and agarose gel. The width of the separation gel 52 can be, for example, a length that allows separation of a sample of 10 to 12 lanes.

  In the present embodiment, a configuration in which the separation gel 52 is filled in the separation unit 50 is employed. However, a configuration in which a number of ultrafine columns called nanopillars are provided between the insulating plate 51 and the insulating plate 53 is also possible. Can be adopted.

  The first opening 50a of the separation part 50 is formed of a conductive porous material (for example, a hydrophilic PVDF (Polyvinylidene difluoride) film, a hydrophilic PTFE (Polytetrafluoroethylene) film, etc.) including the periphery thereof. It may be covered with a covering portion. Accordingly, when the transfer film 1 is in contact with or pressed against the first opening 50a (when no distance is provided between the first opening 50a and the transfer film 1), the transfer film 1 is transferred when the transfer film 1 is conveyed. The frictional resistance and damage received from the separation part 50 and the separation gel 52 can be reduced.

  In addition, since the separation part 50 stands in a substantially vertical direction, the amount of sample introduction can be increased as compared with a configuration in which the separation part 50 is installed in a substantially horizontal direction. This is because it is difficult to change the depth of the well provided in the separation gel in the horizontal electrophoresis apparatus, but the depth of the well can be easily changed in the vertical electrophoresis apparatus. This is because the amount can be easily increased.

(Transfer film 1)
The transfer film 1 is preferably a sample adsorbing / holding body that allows the sample separated by the separation gel 52 to be stably stored for a long period of time and further facilitates subsequent analysis. The material of the transfer film 1 is preferably a material having high strength and high sample binding ability (weight that can be adsorbed per unit area). As the transfer film 1, a PVDF film or the like is suitable when the sample is a protein. Note that the PVDF membrane is preferably hydrophilized in advance using methanol or the like. In addition to this, membranes conventionally used for protein, DNA and nucleic acid adsorption such as nitrocellulose membrane or nylon membrane can also be used.

  The sample that can be separated and adsorbed in the biomolecule analyzer 100 is not limited to these, but is a preparation from a biological material (for example, a living organism, body fluid, cell line, tissue culture, or tissue fragment), or And commercially available reagents. For example, a polypeptide or polynucleotide is mentioned.

  The transfer film 1 is used in a state where it is immersed in a buffer solution in the anode buffer tank 30.

  In this embodiment, the transfer film 1 has a length that is used for one electrophoresis / transfer, in other words, a distance that moves in the anode buffer tank 30 in one electrophoresis / transfer. If you do. By configuring the transfer film 1 in this way, an operation of cutting the transfer film 1 is not required for each electrophoresis / transfer, and the usability of the biomolecule analyzer 100 can be improved. Further, the lateral width of the transfer film 1 may be a length corresponding to the lateral width of the separation gel 52.

(Arm part)
In the present embodiment, the transfer film 1 is used in a state of being held by the arm portion for the movement and the maintenance of the relative position with the first opening 50a. In the present embodiment, the arm portion includes clamps 20 and 21, a clamp frame 22, a carrier 23, and a guide pole 66 that are a series of connected members. Of these, the structure composed of the clamps 20 and 21 and the clamp frame 22 may be referred to as an adjuster. The adjuster is disposed inside the side wall of the anode buffer tank 30. Also,
FIG. 3 is a perspective view showing a schematic structure of the adjuster. As shown in FIG. 3, the clamp 20 includes a lower portion 20a and an upper portion 20b, and the clamp 21 includes a lower portion 21a and an upper portion 21b.

  As shown in FIG. 3B, the space between the lower portion 20a and the upper portion 20b and the space between the lower portion 21a and the upper portion 21b are opened. In this state, an end (first end) in the moving direction of the transfer film 1 is sandwiched between the lower portion 20a and the upper portion 20b, and an end in the moving direction of the transfer film 1 is interposed between the lower portion 21a and the upper portion 21b. After sandwiching the part (second end part), as shown in FIG. 3A, the clamp is performed by closing between the lower part 20a and the upper part 20b and between the lower part 21a and the upper part 21b. The end of the transfer film 1 in the moving direction can be fixed by 20, and the end of the transfer film 1 in the moving direction can be fixed by the clamp 21. As a result, the arm portion can hold the transfer film 1. The clamps 20 and 21 may be provided with a lock for fixing in the closed state.

  The clamp frame 22 is a shaft member that connects the clamps 20 and 21, and connects the clamps 20 and 21 with a predetermined distance therebetween. Thereby, when both ends of the transfer film 1 are fixed by the clamps 20 and 21, the transfer film 1 can be stretched without loosening along the moving direction. Thereby, it can suppress that a transcription | transfer result shakes by the looseness of the transfer film 1, and can improve a measurement sensitivity. Further, the tension applied to the transfer film 1 conveyed along with the movement of the clamp 20 can be made constant. Therefore, the sample can be more suitably transferred to the transfer film 1 without unevenness. However, the structure which abbreviate | omits the clamp 21 and the clamp frame 22 may be sufficient.

  The clamp frame 22 is disposed at a position sandwiching the transfer film 1 from the side in the movement direction, and thereby, the front surface (surface facing the first opening 50a) and the back surface (opposite to the first opening 50a) of the transfer film 1. It can be avoided that the clamp frame 22 overlaps the side surface. Accordingly, it is possible to prevent the clamp frame 22 from inhibiting the transfer from the separation gel 52 to the transfer film 1 and the contact of other members with the back surface of the transfer film 1 (details will be described later).

  The guide pole 66 is a shaft member that is connected to a drive unit (shaft holder 65) described later and is disposed so as to pass outside the side wall of the anode buffer tank 30. The carrier 23 is a member that is connected to the guide pole 66, goes around the upper end of the side wall of the anode buffer tank 30, and is connected to the clamp 20.

  As described above, the arm portion passes from the position connected to the drive portion to the outside of the side wall of the anode buffer tank 30, wraps around the upper end of the side wall, and is connected to the inside of the side wall.

  Although not limiting the present invention, in this embodiment, the guide pole 66 extends outside the side wall of the anode buffer tank 30 to a position aligned with the upper end of the side wall. The carrier 23 is fitted to the guide pole 66 and extends inside the side wall across the upper end of the side wall of the anode buffer tank 30.

  With this configuration, the carrier 23 can be easily attached to and detached from the drive unit. The guide pole 66 is disposed outside the side wall of the anode buffer tank 30, and various operations such as removal of the anode buffer tank 30 (details will be described in Embodiment 2) and electrode setting are performed as necessary. Will not get in the way. Therefore, various operations can be successfully performed by appropriately removing the carrier 23.

(Drive part)
The drive unit drives the arm unit in a substantially horizontal direction, and in the present embodiment, is constituted by a motor 62, a ball screw 63, a guide shaft 64, and a shaft holder 65.

  The motor 62 rotates the ball screw 63. As the motor 62, a motor whose speed can be changed may be used, or a motor having a fixed speed may be used in combination with a gear. The ball screw 63 penetrates the shaft holder 65 and is screwed into the shaft holder 65. The guide shaft 64 passes through the shaft holder 65, and the shaft holder 65 is configured to be movable along the guide shaft 64. Then, when the motor 62 rotates the ball screw 63, the shaft holder 65 is driven in the X direction (substantially horizontal direction) in the figure. The shaft holder 65 is connected to the arm portion (guide pole 66), and thus, the drive portion can drive the arm portion in the X direction (substantially horizontal direction) in the drawing. Since the arm portion holds the transfer film 1, the transfer film 1 moves in the X direction (substantially horizontal direction) in the figure.

  However, the present invention is not limited to this, and the driving unit may be configured by another driving mechanism (for example, a belt, a gear, or the like) as long as the arm unit can be driven in a substantially horizontal direction. Good.

  The drive unit is provided under the anode buffer tank 30. Accordingly, it is possible to prevent the buffer solution scattered from the anode buffer tank 86 from deteriorating the durability of the driving unit and the driving unit from interfering with various operations on the biomolecule analyzer 100.

(Control part)
The control unit 68 is a control panel that performs various controls of the biomolecule analyzer 100 (control of the position of the arm unit, control of current and voltage applied to the anode 32 and the cathode 41, etc.). The control unit 68 may include a button and a switch for receiving an input from the user, a lamp for notifying the user of an operation state, a display unit, and the like.

(Sample electrophoresis and transcription)
Next, the flow of electrophoresis and transfer of the sample in the biomolecule analyzer 100 will be described with reference to FIG. FIG. 4 is a cross-sectional view for explaining electrophoresis and transfer of a sample in the present embodiment. In FIG. 4, the anode buffer tank 30, the cathode buffer tank 40, etc. are omitted for convenience of explanation.

  As shown in FIG. 4, at the time of sample electrophoresis and transfer, the transfer film 1 is held by the clamps 20 and 21 (adjusters, arm portions) in a state of being arranged at a position facing the first opening 50a. . At this time, the transfer film 1 is supported from the back surface of the transfer film 1 (the side opposite to the separation unit 50) by guides 33 and 34 provided at the bottom of the anode buffer tank 30.

  The guides 33 and 34 are provided at the bottom of the anode buffer tank 30 so as to support the transfer film in the moving path along which the transfer film 1 moves. The longitudinal directions of the guides 33 and 34 are orthogonal to the moving direction (X direction) of the transfer film 1 and are parallel to the longitudinal direction of the first opening 50a.

  Then, the separation film 50 is bent so that the side opposite to the separation part 50 is convex when the separation part 50 (on the first opening 50a side) abuts on the surface of the transfer film 1 (separation part 50 side). It has been. In this way, the transfer film 1 is supported by the guides 33 and 34, and the separation unit 50 presses it and is bent so as to protrude downward (to the side opposite to the separation unit 50). As a result, tension is applied to the transfer film 1, and the transfer film 1 can be brought into close contact with the first opening 50a. Thereby, the transfer from the separation gel 52 to the transfer film 1 can be more suitably performed.

  In particular, the guides 33 and 34 are formed at positions sandwiching the position facing the first opening 50 a at the bottom of the anode buffer tank 30, so that the guides 33 arranged on both sides of the separation unit 50 are formed. The transfer film 1 is supported by 34, and the separation part 50 is pressed down and bent so as to protrude downward (opposite to the separation part). As a result, a uniform tension is applied to the transfer film 1, and the transfer film 1 can be evenly adhered to the first opening 50a. Thereby, the transfer from the separation gel 52 to the transfer film 1 can be more suitably performed.

  More specifically, the tension of the transfer film 1 when transferring a sample from the separation gel 52 to the transfer film 1 is preferably, for example, a tension within a range of 1N or more and 12N or less, and is about 6N. Most preferred. If the tension applied to the transfer film is in the above range, it is possible to transfer the sample from the separation gel 52 to the transfer film 1 with high sensitivity and to prevent the transfer film 1 from being damaged by excessive tension. it can.

  In order to set the tension of the transfer film 1 within the above range, for example, the inclination angle θ from the position in contact with the guides 33 and 34 to the position in contact with the first opening 50a in the transfer film 1 is preferably downward with respect to the horizontal plane It can be suitably realized by setting the angle to 1 ° or more and 60 ° or less, more preferably about 10 °. Since the tension of the transfer film 1 is defined by the tilt angle, the tension of the transfer film 1 can be set to the above-described range by setting the tilt angle within the above-described range.

  As described above, since the clamp frame 22 is disposed at a position sandwiching the transfer film 1 from the side in the movement direction, the guides 33 and 34 do not prevent the transfer film 1 from being supported from the back surface.

  Then, the sample is introduced into the separation gel 52 from the second opening 50 b of the separation unit 50. In addition to the biomolecule to be analyzed, it is preferable to add a visible molecular weight marker for confirming the progress of electrophoresis to the sample.

  In the above state, the sample is separated by electrophoresis. The control unit 68 controls the motor 62 to set the position of the transfer film 1 as a start position, and allows current to flow between the anode 32 and the cathode 41 to start electrophoresis. The value of the current that flows between the anode 32 and the cathode 41 is not particularly limited, but is preferably 50 mA or less, and more preferably 20 mA or more and 30 mA or less. The current value may be controlled to be constant, the voltage may be controlled to be constant, or the current / voltage may be controlled in other manners.

  The transfer film 1 is gradually moved in the X direction (substantially horizontal direction) by driving the arm unit (adjuster) by the driving unit in accordance with the progress of electrophoresis in the separation unit 50. The X direction is a direction orthogonal to the longitudinal direction of the first opening 50a. The moving speed of the transfer film 1 is not particularly limited. For example, it can be a pace that moves 5 to 10 cm in 60 to 120 minutes.

  Then, the position of the sample discharged from the first opening 50a by electrophoresis (separated in the separation gel 52) according to the discharge timing in the transfer film 1 (the first opening 50a at the discharge timing). Adsorbed at the position facing the As a result, the separated sample is transferred to the transfer film 1.

  After the transfer, the transfer film 1 can be collected and used for staining or immune reaction (blocking and antigen-antibody reaction in Western blotting) and the like. Thereafter, a separation pattern of components transferred to the transfer film 1 is detected by a fluorescence detector or the like. Such a fluorescence detector may be incorporated in the biomolecule analysis apparatus 100, and thereby, all the steps of electrophoresis, transcription, and detection can be automated.

  As described above, with the configuration in which the separation unit 50 stands substantially vertically, the separation unit 50 is immersed in at least one buffer solution of the anode buffer tank 30 and the cathode buffer tank 40, and the separation gel 52 is cooled with water. Can do.

  When the biomolecule analyzer 100 is configured in this way, it is necessary to (i) move the transfer film 1 in the anode buffer tank 30, and (ii) transfer the drive unit as in the prior art. If it is intended to be arranged upstream of the membrane 1, the buffer solution scattered from the anode buffer tank 30 may reduce the durability of the drive unit, and the drive unit may hinder various operations on the biomolecule analyzer 100. (Iii) In this embodiment, the drive unit is provided under the anode buffer tank 30 and the shape of the arm portion passes through the outside of the side wall of the anode buffer tank 30 and the upper end of the side wall is By forming a shape that wraps around and is connected to the inside of the side wall, the transfer film 1 is an anode while avoiding a decrease in durability of the drive unit due to the buffer solution and obstruction of various operations by the drive unit. It can be moved successfully in Ffa vessel 30. Accordingly, it is possible to provide a vertical discharge transfer type electrophoresis-transfer apparatus including a suitable transfer film transport mechanism.

[Embodiment 2]
It will be as follows if other embodiment of this invention is described based on FIGS. For convenience of explanation, members having the same functions as those described in the embodiment are given the same reference numerals, and descriptions thereof are omitted.

In the biomolecule analyzer 100, the anode buffer tank 30, the cathode buffer tank 40, and the separation unit 50 may be detachable from the biomolecule analyzer 100. Thereby, since the anode buffer tank 30, the cathode buffer tank 40, and the separation part 50 can be removed and cleaned, the first buffer tank can be easily cleaned without attaching a cleaning solution or the like to the drive unit. .

  FIG. 5 is a perspective view showing a state where the separation unit 50 is removed from the biomolecule analyzer 100. Since the separation unit 50 is fixed to the cathode buffer tank 40 by a lock 42, it can be easily removed by releasing the lock 42. The separation unit 50 only needs to be detachably attached to the cathode buffer tank 40, and the method is not limited to the method using the lock 42.

  FIG. 6 is a perspective view showing a state in which the cathode buffer tank 40 is further removed from the biomolecule analyzer 100. The cathode buffer tank 40 is not particularly limited, but may be detachably fixed to the anode buffer tank 30 with, for example, a screw, a lock, or the like.

  FIG. 7 is a perspective view showing a state in which the carrier 23 is further removed from the biomolecule analyzer 100. Since the carrier 23 is coupled to the guide pole 66 and the clamp 20 by fitting to the guide pole 66 and the clamp 20, respectively, the carrier 23 can be easily removed. Note that the carrier 23 only needs to be removable from at least the drive unit. For example, the guide pole 66 may be separable from the shaft holder 65.

  FIG. 8 is a perspective view showing a state where the anode buffer tank 30 is further removed from the biomolecule analyzer 100. As described above, in the present embodiment, since the carrier 23 that is a portion that goes around the upper end of the side wall of the anode buffer tank 30 in the arm portion can be separated from the drive unit, the anode buffer tank 30 can be easily removed. it can. Although the anode buffer tank 30 is not specifically limited, For example, what is necessary is just to be detachably fixed to the table 31 by fitting.

[Embodiment 3]
The following will describe another embodiment of the present invention with reference to FIG. For convenience of explanation, members having the same functions as those described in the embodiment are given the same reference numerals, and descriptions thereof are omitted.

  FIG. 9 is a perspective view showing a schematic configuration of the biomolecule analyzer 100 according to the present embodiment. As shown in FIG. 9, in this embodiment, the anode buffer tank 30, the cathode buffer tank 40, and the separation unit 50 are configured so that the whole or a part thereof is transparent using, for example, transparent resin, glass, or the like. Has been. Further, the lid that covers the whole during the above-described operation is also configured to be transparent. Preferably, the arm portions (clamps 20 and 21, clamp frame 22, carrier 23 and guide pole 66) may also be configured so that the whole or part thereof is transparent. Thereby, the state of the separation gel 52 and the transfer film 1 can be observed during the operation of the apparatus. Thereby, for example, the movement of the visible marker can be confirmed with the naked eye.

[Embodiment 4]
The following will describe another embodiment of the present invention with reference to FIG. For convenience of explanation, members having the same functions as those described in the embodiment are given the same reference numerals, and descriptions thereof are omitted. This embodiment is different from the first embodiment in the configuration of the adjuster, and the other configurations are the same as those in the first embodiment. Hereinafter, the difference in the configuration of the adjuster will be described.

  FIG. 10A is a cross-sectional view showing the configuration of the adjuster in the first embodiment, and FIG. 10B is a cross-sectional view showing the configuration of the adjuster in the present embodiment. Both adjusters fix a clamp (first fixing portion) 20 that fixes the front end (first end) of the transfer film 1 in the moving direction, and a rear end (second end) of the transfer film 1 in the moving direction. A clamp (second fixing portion) 21, and a clamp frame (connecting portion) 22 that connects the clamp 20 and the clamp 21.

  Here, in the adjuster according to the present embodiment, as shown in FIG. 10B, a portion on the clamp 21 side in the clamp frame 22, the insertion portion 22 a having a small diameter, is inserted into the clamp 21. It is inserted. Thus, the clamp 21 is configured to be slidable along the clamp frame 22. Further, an elastic body 22b is provided at a position sandwiched between the clamp frame 22 and the clamp 21, and the clamp 20 connected to the clamp frame 22 and the clamp 21 face each other by the elastic force of the elastic body 22b. It is energized in the direction to do. Here, for example, in a state where the clamp 20 and the clamp 21 are close to each other against the elastic force, both ends of the transfer film 1 are fixed to the clamps 20 and 21, respectively, and the clamp 20 and the clamp 21 are close to each other. If released from the above, both ends of the transfer film 1 are pulled in directions opposite to each other, and a constant tension can be applied to the transfer film 1 so that the transfer film 1 is stretched. If the transfer film 1 is in a relaxed state, when the transfer film 1 is moved, the gap between the transfer film 1 and the first opening 50a may become large and the transfer result may be blurred. In this case, since the transfer film 1 can be stretched, a good transfer result can be obtained.

  In particular, as shown in FIG. 4, the transfer film 1 is supported by the guides 33 and 34, and the separation part 50 is pressed down and bent so as to protrude downward (opposite to the separation part 50). Since the transfer film 1 is maintained in the tensioned state by the elastic body 22b, the transfer film 1 is pressed against the first opening 50a in an attempt to return to the unfolded state. Thereby, the transfer film 1 can be pressed against the first opening 50a so that a good transfer result can be obtained.

  The material and arrangement of the elastic body 22b are not particularly limited as long as the elastic body 22b urges the clamps 20 and 21 in the direction facing away from each other by its elastic force, but is made of a material that does not cause electrolysis. Or coated with a material that does not cause electrolysis. For example, the elastic body 22b may be a spring made of a resin that does not cause electrolysis or a metal coated with a resin. The elastic body 22b may also be an elastic body such as sponge or rubber.

  The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention. Furthermore, a new technical feature can be formed by combining the technical means disclosed in each embodiment.

  The present invention can be used in the field of manufacturing analytical instruments such as biomolecules and the field of analysis of biomolecules.

1 Transfer film 20 Clamp (arm part, first fixing part)
21 Clamp (arm part, second fixing part)
22 Clamp frame (arm part, connecting part)
22b Elastic body 23 Carrier (arm part, part that goes around the upper end of the side wall, second part)
30 Anode buffer tank (first buffer tank)
31 Table 32 Anode (first electrode)
34/35 Guide (support member)
40 Cathode buffer tank (second buffer tank)
41 Cathode (second electrode)
42 Lock 50 Separating part 50a First opening 50b Second opening 51/53 Insulating plate 52 Separation gel (separation medium)
62 Motor (drive unit)
63 Ball screw (drive unit)
64 Guide shaft (drive unit)
65 Shaft holder (drive unit)
66 Guide pole (arm part, first part)
68 Control Unit 100 Biomolecule Analyzer

Claims (12)

First buffer tank,
A second buffer tank disposed above the first buffer tank;
A separation unit in which a separation medium is stored, has a first opening opened in the first buffer solution tank and a second opening opened in the second buffer solution tank, and stands up in a substantially vertical direction,
An arm unit for holding a transfer film disposed at a position facing the first opening; and a drive unit that is provided under the first buffer solution tank and drives the arm unit in a substantially horizontal direction.
The biomolecule analyzer is characterized in that the arm portion is connected to the driving portion and is driven so as to be transferred to the transfer film at a timing when the sample separated from the first opening is discharged.
The transfer film has one end front in the movement direction by driving the arm portion and the other end rear in the movement direction,
The arm portion biases one fixing portion for fixing one end portion, the other fixing portion for fixing the other end portion, and the one fixing portion and the other fixing portion in a direction facing away from each other. The biomolecule analyzer according to claim 1, further comprising an elastic body. A support member for supporting the transfer film from the side opposite to the separation part in the transfer film is provided at the bottom of the first buffer tank,
The biomolecule analyzer according to claim 2, wherein the transfer film is bent by the separation unit so that the side opposite to the separation unit is convex.   4. The biomolecule analyzer according to claim 3, wherein the support member is formed at a position sandwiching a pair of positions facing the first opening at the bottom. The transfer film has one end front in the movement direction by driving the arm portion and the other end rear in the movement direction,
The arm portion includes one fixing portion that fixes one end portion, the other fixing portion that fixes the other end portion, and a connecting portion that connects the one fixing portion and the other fixing portion with a predetermined distance therebetween. The biomolecule analyzer according to claim 1, further comprising:   6. The biomolecule analyzer according to claim 5, wherein the connecting portion is disposed at a position sandwiching the transfer film from the side in the moving direction. A support member for supporting the transfer film from the side opposite to the separation part in the transfer film is provided at the bottom of the first buffer tank,
The biomolecule analyzer according to any one of claims 1, 5, and 6, wherein the transfer film is bent by the separation portion so that the opposite side to the separation portion is convex. .   The biomolecule analyzer according to claim 7, wherein the support member is formed at a position sandwiching a pair of positions facing the first opening at the bottom.   The living body according to claim 8, wherein an inclination angle of the transfer film from a position in contact with the support member to a position in contact with the first opening is 1 ° or more and 60 ° or less downward with respect to a horizontal plane. Molecular analyzer.   The biomolecule analyzer according to any one of claims 1 to 9, wherein the first buffer tank, the second buffer tank, and the separation unit are transparent. The first electrode is disposed in the first buffer tank,
A second electrode is disposed in the second buffer solution tank,
The biomolecule analyzer according to claim 1, wherein the transfer film is disposed so as to be inserted into the first opening and the first electrode. The separation part is detachably attached to the second buffer solution tank,
The biomolecule analyzer according to any one of claims 1 to 11, wherein the second buffer solution tank is detachably attached to the first buffer solution tank.

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