JP2016156704A - Biomolecule analysis device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a biomolecule analysis device which has such a structure that a separation part is vertically arranged and which prevents bubbles generated from an electrode from adversely affecting a contact portion of a transfer film with the separation part.SOLUTION: An insulating electrode cover 35 for releasing bubbles generated from an anode 32 is arranged between the anode 32 and a discharge part 50a of an electrophoresis gel chip 50. The electrode cover 35 is hung down to a first recess 30a provided to a bottom of an anode buffer tank 30.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a biomolecule analyzer for analyzing biomolecules such as proteins.

  In proteome analysis, which plays a central role in post-genomic research, the combination of two-dimensional electrophoresis (2DE) and Western blotting is known as an excellent separation analysis technique. 2DE can separate a proteome into a plurality of components (proteins) with high resolution using various separation media based on two independent physical properties (charge and molecular weight) inherent to proteins. When the protein is further analyzed using the result of separation by 2DE, it is preferable to immobilize a plurality of proteins contained in the separation medium on the transfer membrane by Western blotting. This is because the protein immobilized on the transfer membrane can be stored stably over a long period of time and is easy to analyze. In particular, Western blotting is an indispensable step when comprehensively examining multiple biological characteristics of proteins such as increase / decrease in protein expression and presence / absence of post-translational modification using separation results by 2DE. I can say that.

  The conventional Western blotting method has a problem in that bubbles generated from the anode adhere to the contact portion between the transfer membrane and the separation medium, thereby blurring or distorting the protein pattern immobilized on the transfer membrane. Regarding this problem, Patent Document 1 discloses a device using an anode covered with a cylindrical porous material. The advantage of this device is disclosed in Patent Document 1 as follows. Since the bubbles generated from the anode increase in size when passing through the porous material, the bubbles easily rise to the water surface. Thereby, since it is possible to prevent the minute bubbles from being mixed into the buffer, it is possible to eliminate the adverse effect of the bubbles on the contact portion.

US Pat. No. 5,916,429 (issued June 29, 1999) US Pat. No. 5,234,559 (issued August 10, 1993) Special table hei 9-501774 (announced February 18, 1997)

  In the technique of Patent Document 1, generated bubbles are discharged together from the upper part of the buffer tank, which is effective when the separation medium is installed horizontally. However, when the separation medium is installed vertically, the anode is arranged at the bottom of the buffer tank, so that the bubbles generated from the anode head toward the contact point between the transfer membrane and the separation medium before reaching the water surface, and as a result, The problem still remains that air bubbles adversely affect the contact area.

  The present invention has been made to solve the above-described problems of the present invention. An object of the present invention is to provide a biomolecule analyzer in which the separation unit is arranged vertically, and bubbles generated from the electrodes do not adversely affect the contact portion between the transfer film and the separation unit. .

In order to solve the above problems, a biomolecule analyzer according to an aspect of the present invention is provided.
A buffer bath;
A transfer film disposed in the buffer solution tank;
A transport unit that transports the transfer film along a predetermined transport direction;
A separation unit that vertically contacts the transfer film and is vertically installed, separates the specimen by electrophoresis, and discharges the separated specimen to the transfer film;
An electrode disposed at a position away from the contact portion between the separation part and the transfer film in the transport direction by a certain distance;
An insulating electrode cover disposed between the electrode and the contact portion;
A first recess is provided at a position sandwiched between the electrode and the contact portion at the bottom of the buffer tank,
The electrode cover is characterized by hanging toward the first recess.

  According to one aspect of the present invention, there is provided a biomolecule analyzer having a configuration in which a separation unit is arranged vertically, and bubbles generated from an electrode do not adversely affect the contact portion between the transfer film and the separation unit. There is an effect that can be done.

It is a figure which shows arrangement | positioning of the anode cover in the biomolecule analyzer which concerns on Embodiment 1 of this invention. It is a figure explaining the outline of the transfer film with a frame concerning one embodiment of the present invention. It is a figure explaining the outline of the biomolecule analyzer which concerns on Embodiment 1 of this invention. It is a figure explaining the outline of the electrode cover with a supporting member which concerns on the modification 1 of this invention. It is a figure explaining the outline of the biomolecule analyzer which concerns on Embodiment 2 of this invention. It is a figure explaining the outline of the biomolecule analyzer which concerns on Embodiment 3 of this invention. It is a figure explaining the outline of the biomolecule analyzer which concerns on Embodiment 4 of this invention.

Embodiment 1
A first embodiment according to the present invention will be described below with reference to FIGS.

(Transfer film with frame 10)
First, a framed transfer film used in the biomolecule analyzer according to the present embodiment will be described below with reference to FIG. FIG. 2A is a diagram for explaining the outline of the framed transfer film 10. FIG. 2B is a cross-sectional view taken along line A and line B in FIG.

  As shown in FIG. 2 (a), the framed transfer film 10 includes a pair of frames (frame members) in which one side (first side) of the transfer film 1 and one side (second side) opposite to the one side are arranged. 2 individually supported.

  As shown in the cross-sectional views taken along lines A and B in FIG. 2B, the frame-attached transfer film 10 has an opening-like fitting portion (through the frame 2) as the fitting portion 3 in the pair of frames 2. More preferably, a through hole is provided. Each fitting part 3 can be fitted into a corresponding convex fitting part. Thereby, the transfer film 10 with a frame and the biomolecule analyzer can be fixed by the fitting portion 3.

  The transfer film 10 with a frame can stretch the transfer film 1 without loosening by fixing the pair of frames 2 in parallel and separating them from each other. In addition, since the transfer film 1 that has been cut into a predetermined shape can be fixed by the frame 2, the transfer film 10 with the frame is attached to the biomolecule analyzer while applying a simple and appropriate tension to the transfer film 1. be able to. For this reason, it is possible to prevent the distortion of the blot when it is fixed to the biomolecule analyzer and the specimen is transferred by discharge transfer.

  Further, in the transfer film with frame 10, the frame 2 can be used as the weight of the transfer film 1. For this reason, it is possible to prevent the transfer film 1 from moving in the reagent in a reagent tank such as a shaker and causing unevenness in antibody reaction. Furthermore, since the transfer film 10 with a frame has a simple configuration including the transfer film 1 and a pair of frames 2, for example, the transfer film is flattened and fixed by fixing the entire outer periphery with a frame. In addition, it is not bulky as compared with the case where the transfer film is spread and fixed by a curved frame, and for example, the amount of antibody used when performing Western blotting can be reduced.

(Transfer film 1)
The transfer film 1 is a film for adsorbing and holding a biomolecule sample (analyte) separated by the separation unit of the biomolecule analyzer. Here, the transfer film 1 is a biomolecule sample adsorbing / holding body that enables the biomolecule sample (analyte) separated by the separation unit to be stably stored for a long period of time and further facilitates subsequent analysis. Is preferred. 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 polyvinylidene fluoride (PVDF) film is suitable when the sample is protein. Note that the PVDF membrane is preferably hydrophilized in advance using methanol or the like. In addition to this, it is also possible to use a membrane conventionally used for adsorption of protein, DNA and nucleic acid, such as a nitrocellulose membrane or a nylon membrane.

  Biomolecule samples that can be separated and adsorbed in a biomolecule analyzer can include, but are not limited to, proteins, DNA and RNA. For example, preparations from biological materials (eg, living organisms, body fluids, cell lines, tissue cultures, or tissue fragments), commercially available reagents, and the like are also included in examples of samples. Furthermore, a polypeptide or polynucleotide is a kind of sample.

(Frame 2)
Each of the pair of frames 2 is preferably longer than the length of one side of the transfer film 1 to which the length of the frame 2 is fixed. The frame 2 is preferably made of an insulating material. As the insulating material, resins such as polymethyl methacrylate (acrylic), polystyrene, polyethylene, polypropylene, polyethylene terephthalate (PET), polyacetal (POM), polyetheretherketone (PEEK), or glass can be used.

  The frame 2 is more preferably subjected to a hydrophilic treatment on the surface. For example, a coating layer may be provided on the surface of the frame 2 made of the above material. Thereby, it is possible to prevent a specimen such as a protein discharged from the discharge portion of the separation portion from adhering to the surface of the frame 2, and to prevent the frame 2 from being contaminated.

  The water contact angle on the surface of the frame 2 is preferably 90 ° or less, and more preferably 60 ° or less. By setting the water contact angle on the surface of the frame 2 to 90 ° or less, the frame 2 can be suitably prevented from being contaminated by the specimen.

(Biomolecular analyzer 100)
Next, a biomolecule analyzer 100 according to an embodiment of the present invention will be described in detail with reference to FIG. FIG. 3 is a diagram for explaining the outline of the biomolecule analyzer 100 according to one embodiment of the present invention. As shown in this figure, the biomolecule analyzer 100 includes a clamp 20, an anode buffer tank 30 (buffer tank), an anode stage 31, a cathode buffer tank 40, an electrophoresis gel chip 50 (separation unit), and a transport unit. In addition, a first recess 30 a is provided at the bottom of the anode buffer tank 30.

  In the biomolecule analyzer 100, the anode buffer tank 30 is fixed to the anode stage 31 in a detachable state. A clamp 20 is disposed in the anode buffer tank 30, and the transfer film 10 with the frame is fixed to the clamp 20 in the anode buffer tank 30. The cathode buffer tank 40 is fixed to the anode buffer tank 30 in a detachable state. The electrophoresis gel chip 50 is arranged in the biomolecule analyzer 100 so that one end of both ends facing each other is located in the anode buffer tank 30 and the other end is located in the cathode buffer tank 40. .

  The outline of the biomolecule analyzer 100 is as follows. The electrophoresis gel chip 50 separates the sample introduced into the separation gel 52 by electrophoresis, and discharges the separated sample to the transfer film 1 through the discharge part 50 a at one end of the electrophoresis gel chip 50. The transport unit transports the transfer film 1 in the transport direction X (a direction from one side (first side) where the frame 2 is provided in the frame-equipped transfer film 10 toward the other side (second side) where the frame 2 is provided). Transport to. Thereby, the discharged specimen is adsorbed to a position according to the discharge timing in the transfer film 1 (a position facing the electrophoresis gel chip 50 at the discharge timing). Thereby, the separated specimen is transferred to the transfer film 1.

(Clamp 20)
As shown in FIG. 3, the clamp 20 includes front clamps 20 a and 20 b, rear clamps 21 a and 21 b, and a clamp frame 22. The front clamps 20a and 20b are arranged on the conveyance end point side when the transfer film 1 is conveyed, while the rear clamps 21a and 21b are arranged on the conveyance start point side.

  In the clamp 20, the front clamp 20a and the front clamp 20b are fixed so as to be released by a jig. Fitting portions (not shown) are provided at two places on the front clamp 20a, and fittings provided at two places on one frame 2 shown in FIG. Each part 3 is fitted. Thereafter, one frame 2 is sandwiched and fixed by the front clamp 20a and the front clamp 20b.

  Similarly, in the clamp 20, the rear clamp 21a and the rear clamp 21b are fixed so as to be released by a jig. Fitting portions (not shown) are provided at two locations in the rear clamp 21a, and fitting portions provided at two locations on the other frame 2 shown in FIG. 2 (a) with respect to these fitting portions. 3 is fitted. Thereafter, the other frame 2 is sandwiched and fixed by the rear clamp 21a and the rear clamp 21b.

  The clamp frame 22 fixes the front clamp 20a and the rear clamp 21a in a state of being separated by a certain distance. For this reason, when the transfer film 10 with a frame is fixed by the clamp 20, the transfer film 1 is fixed in a stretched state without loosening. Further, the clamp frame 22 fixes the front clamp 20a and the rear clamp 21a from the side of the transfer film 1 in the transport direction (outside of the two sides not supported by the frame 2). Thus, when the transfer film 10 with the frame is fixed to the biomolecule analyzer 100 via the clamp 20, the clamp frame 22 is prevented from contacting the electrophoresis gel chip 50 and the guides 33 and 34 in the transfer path of the transfer film 1. The transfer film 10 with a frame can be disposed.

  The carrier 23 is provided on the front clamp 20b. When the clamp 20 is attached to the inside of the anode buffer tank 30, the clamp 20 can be fixed to the guide pole 66 disposed outside the anode buffer tank 30 through the carrier 23 in a detachable state.

(Anode buffer tank 30)
In FIG. 3, the anode buffer tank (buffer tank) 30 is indicated by a dotted line. As shown in FIG. 3, the anode buffer tank 30 includes an anode (electrode) 32, guides (support members) 33 and 34, and an anode cover 35 (electrode cover). A recess 30 a is provided at the bottom of the anode buffer tank 30.

  The anode buffer tank 30 is filled with an anode buffer. Examples of the anode buffer include Tris / glycine buffer solution, acetate buffer solution, sodium carbonate buffer solution, CAPS buffer solution, Tris / boric acid / EDTA buffer solution, Tris / acetic acid / EDTA buffer solution, MOPS, phosphate buffer. Or a buffer such as Tris / Tricine buffer. The transfer film 10 with the frame fixed to the clamp 20 is installed in an anode buffer filled in the anode buffer tank 30.

  The anode 32 is an elongated rod-like electrode made of a platinum wire or the like. The anode 32 is provided at the bottom of the anode buffer tank 30 so that the length direction thereof is perpendicular to the transport direction X of the framed transfer film 10. The anode 32 is not located directly under the discharge part 50a in the electrophoresis gel chip 50, but is disposed at a position away from the discharge part 50a in the transport direction X by a certain distance. This position is a position where a voltage can be applied between the anode 32 and the cathode 41 from the back surface of the transfer film 1 on the side facing the electrophoresis gel chip 50 when the framed transfer film 10 is installed.

  The guides 33 and 34 support a pair of positions sandwiching from the front and rear in the transport direction from the opposite side of the transfer gel 1 to the electrophoresis gel chip 50, respectively. It is a supporting member to do. The guides 33 and 34 are provided at the bottom of the anode buffer tank 30 on the conveyance path through which the frame-attached transfer film 10 is conveyed. The guides 33 and 34 are arranged so that their height directions are parallel to the in-plane direction of the electrophoresis gel chip 50 and perpendicular to the transport direction X in which the framed transfer film 10 is transported. Yes. As a result, the guides 33 and 34 allow the transfer film 1 to be parallel to the length direction of the end of the electrophoresis gel chip 50 on the discharge part 50a side from the back surface of the transfer film 1 facing the electrophoresis gel chip 50. To support.

(Anode cover 35)
The anode cover 35 is provided in contact with the anode 32 or apart from the anode 32 between the anode 32 and a contact portion between the transfer film 1 and the electrophoresis gel chip 50. As will be described in detail later, since the biomolecule analyzer 100 includes the anode cover 35, air bubbles generated from the electrode from the anode 32 can escape to the upper part of the anode buffer tank 30. It is possible to prevent bubbles from adversely affecting the contact portion with the electrophoresis gel chip 50.

  The anode cover 35 is made of an insulating material. As the insulating material, resins such as polymethyl methacrylate (acrylic), polystyrene, polyethylene, polypropylene, polyethylene terephthalate (PET), polyacetal (POM), polyetheretherketone (PEEK), or glass can be used.

  The anode cover 35 is more preferably subjected to hydrophilic treatment on the surface. For example, a coating layer may be provided on the surface of the frame 2 made of the above material, for example. Thereby, bubbles generated from the anode 32 can be easily released along the surface of the anode cover 35 to the upper part of the anode buffer tank 30.

  The contact angle with water on the surface of the anode cover 35 is preferably 90 ° or less, and more preferably 60 ° or less. By setting the water contact angle on the surface of the anode cover 35 to 90 ° or less, bubbles can be more easily released along the surface of the anode cover 35.

(Cathode buffer tank 40)
As shown in FIG. 3, the cathode buffer tank 40 includes a cathode 41 and a lock 42. The cathode 41 is an elongated rod-like electrode composed of a platinum wire or the like. The cathode 41 is arranged on the upper side (directly above the separation gel 52) inside the cathode buffer tank 40 so that the length direction thereof is perpendicular to the transfer direction of the transfer film 1. That is, the length direction of the cathode 41 is parallel to the length direction of the anode 32.

  The cathode buffer tank 40 is filled with a cathode buffer. As the cathode buffer, a buffer similar to the anode buffer can be used.

  In the cathode buffer tank 40, the electrophoresis gel chip 50 is fixed by a lock 42. At this time, the end of the electrophoresis gel chip 50 opposite to the discharge part 50 a is immersed in the cathode buffer filled in the cathode buffer tank 40. On the other hand, the end of the electrophoresis gel chip 50 on the discharge part 50 a side is immersed in an anode buffer filled in the anode buffer tank 30.

  As shown in FIG. 3, the end of the separation gel 52 opposite to the discharge part 50 a side faces the cathode 41. On the other hand, the end of the separation gel 52 on the discharge part 50 a side does not face the anode 32. As described above, the end of the separation gel 52 on the discharge portion 50 a side is separated from the anode 32 by a certain distance in the transfer direction of the transfer film 1.

(Electrophoresis gel chip 50)
As shown in FIG. 3, the electrophoresis gel chip 50 includes an insulating plate 51, a separation gel 52, and an insulating plate 53. The insulating plate 51 and the insulating plate 53 are formed of a plate made of an insulator such as glass and acrylic, for example. A separation gel 52 is formed between the insulating plate 51 and the insulating plate 53.

  The separation gel 52 is a gel for separating the introduced biomolecule sample (analyte) according to the molecular weight. Examples of the separation gel 52 include polyacrylamide gels and agarose gels, and it is preferable to use a gel that is suitable for the above-described composition and matched to a buffer solution. The separation gel 52 can be formed by filling the electrophoresis gel chip 50 before attaching the electrophoresis gel chip 50 to the cathode buffer tank 40.

  The electrophoresis gel chip 50 is arranged in the biomolecule analyzer 100 so as to abut on the transfer film 1 perpendicularly. Furthermore, the electrophoresis gel chip 50 is arranged vertically. The discharge portion 50 a of the electrophoresis gel chip 50 is in contact with the surface of the transfer film 1. In the electrophoresis gel chip 50, the biomolecule sample is supplied to the separation gel 52 through the end facing the discharge part 50 a and disposed in the cathode buffer tank 40. After the biomolecule sample is supplied, electrophoresis is performed by applying a voltage between the anode 32 and the cathode 41. As a result, the specimen is transferred to the transfer film 1 through the discharge portion 50a.

(Transport section)
As shown in FIG. 3, the transport unit includes a motor 62, a ball screw 63, a guide shaft 64, a shaft holder 65, and a guide pole 66.

  In the transport unit, the shaft holder 65 can be moved in the transfer direction X along the guide shaft 64 by rotating the ball screw 63 by the motor 62. A guide pole 66 is fixed to the shaft holder 65, and the guide pole 66 supports the carrier 23 provided on the clamp 20 from the outside of the anode buffer tank 30.

  With the above-described configuration, the transfer unit rotates the motor 62, thereby transferring the frame-attached transfer film 10 disposed inside the anode buffer tank 30 via the guide pole 66 disposed outside the anode buffer tank 30. Is moved in the transfer direction X.

(Operation of Biomolecule Analyzer 100)
The operation of the biomolecule analyzer 100 will be described below. First, the transfer film 10 with the frame is fixed by the clamp 20 and placed inside the anode buffer tank 30 filled with the anode buffer. The transfer film 1 of the transfer film with frame 10 is fixed in a state of being supported from below by the guide 33 and the guide 34.

  Thereafter, the cathode buffer tank 40 to which the electrophoresis gel chip 50 is fixed by the lock 42 is fixed to the upper part of the anode buffer tank 30. At this time, the cathode buffer tank 40 is installed in such a state that the electrophoresis gel chip 50 is pressed against the upper side of the transfer film 1. As a result, the transfer film 1 is fixed in a state of being bent (in a valley fold) so as to protrude to the opposite side of the electrophoresis gel chip 50 by contacting the guide 33, the guide 34, and the electrophoresis gel chip 50. .

  Next, by applying a voltage between the anode 32 and the cathode 41, the transfer film 1 of the frame-attached transfer film 10 in the anode buffer transfers the specimen discharged by the electrophoresis gel chip 50 while transferring the sample. With the discharge part of the electrophoresis gel chip 50 being pressed, it is conveyed in the transfer direction X shown in FIG. For this reason, the tension generated when the transfer film 1 is conveyed concentrates on the discharge portion provided at the end of the electrophoresis gel chip 50. That is, the transfer film 1 is conveyed in the transfer direction X while being pressed against the discharge portion of the electrophoresis gel chip 50 by a certain force.

  For this reason, in the transfer film 10 with a frame, it is possible to prevent a gap from being generated between the transfer film 1 and the specimen discharge portion of the electrophoresis gel chip 50 when the transfer film 1 is transported. Therefore, it is possible to suppress the specimen discharged from the discharge portion of the electrophoresis gel chip 50 from diffusing into the anode buffer before being transferred to the transfer film 1. Thereby, the fluctuation of the band of the specimen transferred to the transfer film 1 can be reduced, and the sensitivity of the biomolecule analyzer can be improved.

(Arrangement of anode cover 35)
FIG. 1 is a diagram showing the arrangement of the anode cover 35 in the biomolecule analyzer 100 according to the present embodiment. In this figure, only some members provided in the biomolecule analyzer 100 are illustrated. In the example of FIG. 1, the anode cover 35 has a trapezoid shape whose bottom surface is inclined in the transfer direction of the transfer film 1.

  The anode cover 35 has an end side (end portion) 35 a and an end side 35 b that are orthogonal to the transport direction X and face each other on the bottom surface. In the transport direction X, the end side 35a is closer to the contact point between the transfer film 1 and the electrophoresis gel chip 50 (corresponding to the discharge part 50a) than the end side 35b. In other words, in the transport direction X, the end side 35b is farther from the discharge portion 50a than the end side 35a.

  The bottom surface of the anode cover 35 is inclined such that the end side 35a is closer to the bottom of the anode buffer tank 30 than the end side 35b. In other words, the bottom surface of the anode cover 35 is inclined so that the end side 35b is closer to the water surface of the anode buffer than the end side 35a. The inclination angle is not particularly limited, and may be a fixed angle, or the inclination angle may change depending on the position.

  Further, in the lower part of the contact portion between the transfer film 1 and the electrophoresis gel chip 50, the anode buffer tank 30 has a groove-shaped first recess 30a so that the longitudinal direction thereof is orthogonal to the transport direction of the transfer film 1. Is provided. The first recess 30a has a depth of about 2 to 3 mm.

  An end side 35a of the anode cover 35 hangs down toward the first recess 30a. Preferably, the end side 35a of the anode cover 35 is loosely inserted into the first recess 30a along the longitudinal direction of the first recess 30a. Thus, a bottleneck is formed between the anode cover 35 and the first recess 30a while the end side 35a of the anode cover 35 is disposed at a position lower than the bottom surface of the anode buffer tank 30 in which the anode 32 is disposed. To do.

  Since the bottom surface of the anode cover 35 is inclined as shown in FIG. 1, bubbles generated in the anode 32 can be released in the winding direction Y of the transfer film 1 along the surface of the anode cover 35. Further, since the end side 35a of the anode cover 35 is disposed at a position lower than the bottom of the anode 32, air bubbles generated from the anode 32 pass through the bottleneck formed by the end side 35a and the first recess 30a. There is nothing to do. For this reason, the bubbles escape in a direction away from the discharge part 50a and do not move to the end side 35a side (discharge part 50a side). As a result, bubbles do not adversely affect the contact portion between the transfer film 1 and the electrophoresis gel chip 50.

(Narrowing down electric lines of force)
As described above, the anode 32 is not located directly under the discharge unit 50a, but is disposed at a position away from the discharge unit 50a by a certain distance in the transport direction X. Therefore, if there is no anode cover 35, the lines of electric force generated from the discharge part 50a during electrophoresis are greatly spread. When the lines of electric force are greatly spread, the sample discharged from the discharge part 50a is diffused greatly, and as a result, the separation performance of the sample may be greatly reduced.

  On the other hand, in the present embodiment, as shown in FIG. 1, the edge 35 a of the anode cover 35 is located on the anode 32 side at the contact portion between the electrophoresis gel chip 50 and the transfer film 1 at the bottom of the anode buffer tank 30. It arrange | positions on the 1st recessed part 30a provided so that it might be located in the lower part of an edge part. Since the anode cover 35 is made of an insulating material, the electric lines of force 70 generated from the discharge part 50a are obstructed by the anode cover 35 present in the immediate vicinity of the discharge part 50a and spread toward the anode 32. Absent. Further, the bottleneck formed by the edge 35 a of the anode cover 35 and the first recess 30 a of the anode buffer tank 30 communicates between the anode 32 and the discharge part 50 a, and the discharge part 50 a at the bottom of the anode buffer tank 30. It is open towards. As a result, as shown in FIG. 1, it is possible to generate a narrowed line of electric force 70 from the discharge portion 50a. Thereby, the separation performance of the specimen can be greatly improved.

  Further, the anode cover 35 can be easily removed from the biomolecule analyzer 100. Therefore, the anode cover 35 can be easily cleaned or washed regularly. Furthermore, the anode cover 35 that has deteriorated over time can be replaced with a new one.

[Modification]
The anode cover provided in the biomolecule analyzer according to this embodiment is not limited to the above embodiment. For example, as shown in FIGS. 4A and 4B, the anode cover 35 and the guide 33 may be integrally formed. 4A is a view for explaining the outline of an anode cover with a support member according to a modification of the present invention, and FIG. 4B is a cross-sectional view taken along line A in FIG. 4A. FIG.

  As shown in FIG. 4A, the two support portions 33b of the guide 33 are fixed to both ends of the side surface in the longitudinal direction of the anode cover 35, and the shaft portion 33a is supported by the two support portions 33b. ing.

  As shown in FIG. 4B, the anode cover 35 has a support portion 33b fixed to the side surface on the end side 35b side, and there is no shielding under the shaft portion 33a supported by the support portion 33b. It is open. Further, side walls 35c perpendicular to the bottom surface of the anode cover 35 are provided at both ends in the longitudinal direction of the anode cover 35 along two sides intersecting the end side 35a and the end side 35b (FIG. 4). (B) shows the side wall portion 35c visible in the back).

  In the biomolecule analyzer 100, the guide 33 is disposed so as to support the transfer film 1 conveyed in the conveying direction by the shaft portion 33a, and the anode cover 35 has an end 35a on the concave portion 30a of the anode buffer tank 30. The end side 35b is disposed at a position farther from the end side 35a than the contact portion between the transfer film 1 and the electrophoresis gel chip 50. Further, the side wall portion 35 c in the anode cover 35 is disposed along the transfer direction of the transfer film 1.

  Bubbles generated from the anode 32 flow from the two sides along the conveying direction of the anode cover 35 toward the contact portion between the transfer film 1 and the electrophoresis gel chip 50 by the side wall portion 35c of the anode cover 35. Is prevented. Accordingly, the bubbles generated from the anode 32 move from the end side 35a side of the anode cover 35 toward the end side 35b side along the bottom surface and the side wall portion 35c of the anode cover 35, and the lower portion of the shaft portion 33a in the guide 33 To the upper part of the anode buffer tank 30. Since the end side 35b of the anode cover 35 is disposed at a position far from the contact portion between the transfer film 1 and the electrophoresis gel chip 50, bubbles escaped from the lower portion of the shaft portion 33a in the guide 33 are brought into contact with the contact portion. Not reach.

[Embodiment 2]
A second embodiment according to the present invention will be described below with reference to FIG.

  As shown in FIG. 5A, each member in the biomolecule analyzer 101 is the same as that of the biomolecule analyzer 100 according to the first embodiment. However, in the anode buffer tank 30, the first recess 30b is provided at a position away from the recess 30a by a certain distance in the transport direction, and the anode 32 is disposed inside the second recess 30b. Different from the embodiment.

  The second recess 30b is provided at a position away from the first recess 30a in the transport direction to the same extent as the position where the anode 32 is disposed in the first embodiment.

  As shown in FIG. 5B, by arranging the anode 32 inside the second recess 30b, minute bubbles B generated from the anode 32 spread in the horizontal direction of the anode 32 during electrophoresis. It can be prevented from moving. Further, the minute bubbles B generated from the anode 32 can be aggregated inside the second recess 30b to grow into larger bubbles B. Thereby, the bubble B can be floated more suitably toward the upper part of the 2nd recessed part 30b, and can be moved along the bottom face of the anode cover 35. FIG. Thereby, it is possible to further prevent the minute bubbles B generated from the anode 32 from moving toward the discharge portion 50a. Therefore, the bubble B does not adversely affect the contact portion between the transfer film 1 and the electrophoresis gel chip 50.

  The depth of the second recess 30b may be a depth that allows the anode 32 to be accommodated inside, but is preferably deeper than the height of the anode 32 in order to more suitably aggregate the bubbles.

[Embodiment 3]
A third embodiment of the present invention will be described below with reference to FIG.

  FIG. 6 is a diagram showing a configuration of each anode cover and anode buffer tank in the biomolecule analyzers 102 and 103 according to the present embodiment.

  A biomolecule analyzer 102 shown in FIG. 6A includes an anode cover 36 instead of the anode cover 35. Unlike the bottom surface of the anode cover 35, the bottom surface of the anode cover 36 is provided with a convex portion 36 a that hangs down toward the first concave portion 30 a along the edge on the discharge portion 50 a side. The longitudinal direction of the convex portion 36a is parallel to the longitudinal direction of the first concave portion 30a. In the transfer direction of the transfer film 1, the convex portion 36a is disposed closer to the discharge portion 50a than the anode 32 provided inside the second concave portion 30b.

  The convex portion 36 a is loosely inserted into the first concave portion 30 a, whereby the anode cover 36 and the first concave portion 30 a form a bottleneck on the bottom surface of the anode buffer tank 30.

  In the anode cover 36, since the convex part 36a protrudes toward the inner side of the first concave part 30a, the convex part 36a can be deeply inserted into the inner side of the first concave part 30a. For this reason, when the bubble generated from the anode 32 floats toward the bottom surface of the anode cover 36, the bubble 36a can prevent the bubble from moving toward the discharge portion 50a, and the bubble Can be moved along the bottom surface of the anode cover 36 toward the end side 36b. Accordingly, it is possible to further prevent bubbles from adversely affecting the contact portion between the transfer film 1 and the electrophoresis gel chip 50.

  Further, the bottleneck formed by the convex portion 36a and the first concave portion 30a is opened at the lower portion of the contact portion between the transfer film 1 and the electrophoresis gel chip 50. For this reason, it is possible to generate a narrowed line of electric force from the discharge portion 50a.

  The biomolecule analyzer 103 shown in FIG. 6B includes an anode cover 37 instead of the anode cover 35. Unlike the bottom surface of the anode cover 35, the bottom surface of the anode cover 37 is farther from the discharge part 50 a than the edge 37 a and closer to the discharge part 50 a than the anode 32 in the transfer direction of the transfer film 1. , A convex portion 37a 'is provided. The convex portion 37 a ′ is loosely inserted into the first concave portion 30 a provided at a position facing the convex portion 37 a ′ in the anode buffer tank 30. As a result, a bottleneck is formed by the portion of the bottom surface of the anode cover 37 from the convex portion 37a 'to the end side 37a and a part of the bottom surface of the anode buffer tank 30 including the first concave portion 30a. For this reason, it is possible to prevent bubbles generated from the anode 32 from moving toward the discharge portion 50a by the convex portion 37a 'provided at a position farther from the discharge portion 50a than the end side 37a.

  The bottleneck is opened at the lower part of the discharge part 50a by the part from the convex part 37a 'to the end side 37a and a part of the bottom surface of the anode buffer tank 30 including the first concave part 30a. For this reason, it is possible to generate a narrowed line of electric force from the discharge portion 50a.

[Embodiment 4]
A fourth embodiment of the present invention will be described below with reference to FIG.

  (A) of FIG. 7 is a figure which shows the structure of each anode cover and anode buffer tank in the biomolecule analyzer 104 which concerns on this embodiment.

  A biomolecule analyzer 104 shown in FIG. 7A includes an anode cover 38 instead of the anode cover 36. Further, the bottom of the anode buffer tank 30 is provided with a first recess composed of recesses 30 a and 30 a ′. A recess 30a is provided below the discharge part 50a at the bottom of the anode buffer tank 30, and 30a 'is provided between the recess 30a and the second recess 30b. The recesses 30 a and 30 a ′ are provided in parallel to each other so that the longitudinal direction is orthogonal to the transfer direction of the transfer film 1.

  On the bottom surface of the anode cover 38, a plurality of convex portions 38a and 38a 'are provided in parallel to the longitudinal direction of the concave portions 30a and 30a' instead of the convex portion 36a of the anode cover 36. The convex portion 38a of the anode cover 37 is loosely inserted into the concave portion 30a, and the convex portion 38a 'is loosely inserted into the concave portion 30a'. Thereby, a bottleneck is formed by the convex portions 38 a and 38 a ′ of the anode cover 37 and the concave portions 30 a and 30 a ′ of the anode buffer tank 30.

  As shown in FIG. 7B, in the anode cover 38, the convex portions 38a and 38a 'can be individually loosely inserted toward the inside of the concave portions 30a and 30a'. For this reason, even if the bubble B generated from the anode 32 moves toward the discharge portion 50a beyond the convex portion 38a ′, the bubble B exceeding the convex portion 38a ′ is once floated in the bottleneck, It can accumulate between convex part 38a and convex part 38a ', and can be isolated from the flow of the electrophoresis buffer. Thereby, it is possible to make it difficult for the bubbles B to flow toward the discharge portion 50a due to the flow of the electrophoresis buffer. Therefore, it is possible to further prevent the bubbles B generated from the anode 32 from moving toward the discharge unit 50a.

  Further, since the bottleneck formed by the convex portions 38a and 38a ′ of the anode cover 38 and the first concave portions 30a and 30a ′ of the anode buffer tank 30 is opened at the lower portion of the discharge portion 50a, the discharge portion 50a. From this, it is possible to generate a narrowed electric field line.

[Another embodiment]
The biomolecule analyzer according to the present invention is not limited to the above embodiment. For example, in the biomolecule analyzer according to another embodiment, the shape of the first recess in the side view may be a semicircular shape, a rectangular shape, a V shape (taper), or the like. In addition, when the electrode cover is deflated with the convex portion that hangs down toward the first concave portion, the shape of the convex portion in a side view may be a semicircular shape such as a rectangle or a V-shape (taper). May be. The shape of the electrode cover and the first concave portion is not limited as long as a bottleneck communicating with the contact portion between the separation portion and the transfer film can be formed between the electrode cover and the first concave portion.

  In the biomolecule analyzer according to still another embodiment, in the buffer solution tank, the end side (end portion) of the electrode cover disposed on the first recess side is the buffer solution on the first recess. It is arranged at the same height as the bottom of the tank. If it is said structure, it will prevent that the bubble which generate | occur | produces from an electrode passes above a 1st recessed part, but flows to the contact location of a transfer film and an electrophoresis gel chip, and an electrode cover and 1st A bottleneck can be formed by the recess.

  In the biomolecule analyzer according to another embodiment, the transfer film 1 may not be provided with the frame 2, and the clamp 20 may directly fix the transfer film 1.

[Additional Notes]
A biomolecule analyzer according to aspect 1 of the present invention is provided.
A buffer bath;
A transfer film disposed in the buffer solution tank;
A transport unit that transports the transfer film along a predetermined transport direction;
A separation unit that vertically contacts the transfer film and is vertically installed, separates the specimen by electrophoresis, and discharges the separated specimen to the transfer film;
An electrode disposed at a position away from the contact portion between the separation part and the transfer film in the transport direction by a certain distance;
An insulating electrode cover disposed between the electrode and the contact portion;
A first recess is provided at a position sandwiched between the electrode and the contact portion at the bottom of the buffer tank,
The electric cover hangs down toward the first recess.

  According to said structure, it can prevent by the electrode cover that the bubble which generate | occur | produces from an electrode at the time of electrophoresis moves toward the contact location of a separation part and a transfer film. In addition, since the electrode cover hangs down toward the first recess provided at the bottom of the buffer solution tank, bubbles generated from the electrode exceed the first recess, and the contact between the separation portion and the transfer film It can prevent moving toward the place. As a result, bubbles generated from the electrode do not adversely affect the contact portion between the transfer film and the separation portion.

  In addition, since the contact portion side and the electrode side of the separation portion and the transfer film communicate with each other through a narrow path formed by the insulating electrode cover and the first concave portion, The field lines can be narrowed down.

In the biomolecule analyzer according to aspect 2 of the present invention, in the aspect 1,
The buffer tank is provided with a second recess at a position away from the first recess by a certain distance in the transport direction,
The electrode is disposed in the second recess.

  According to said structure, it can prevent by the 2nd recessed part that the bubble which generate | occur | produces from an electrode at the time of electrophoresis spreads in a horizontal direction. Moreover, minute bubbles generated from the electrode can be agglomerated inside the second concave portion to grow into larger bubbles. For this reason, it is possible to further prevent the bubbles generated from the electrode from moving toward the contact portion between the transfer film and the separation portion through the bottleneck formed by the first recess and the electrode cover.

In the biomolecule analyzer according to aspect 3 of the present invention, in the above aspect 1 or 2,
The electrode cover includes a convex portion that hangs down toward the first concave portion.

  According to said structure, it can further prevent that the bubble which generate | occur | produced from the electrode by the convex part moves toward the contact location of a separation part and a transfer film along the bottom face of an electrode cover.

In the biomolecule analyzer which concerns on aspect 4 of this invention, in the said aspect 3,
The convex portion in the electrode cover is composed of a plurality of convex portions,
The first recess in the buffer tank is composed of a plurality of recesses,
The plurality of convex portions are arranged so as to hang individually in the plurality of concave portions.

  According to said structure, it can prevent further that the bubble which generate | occur | produced from the electrode moves toward the contact part of a separation part and a transfer film along the bottom face of an electrode cover by a some convex part. it can.

In the biomolecule analyzer according to Aspect 5 of the present invention, in any one of Aspects 1 to 4,
The electrode cover is characterized in that an end portion of the electrode cover or the convex portion is loosely inserted into the first concave portion.

  According to the above configuration, it is possible to further prevent bubbles generated from the electrode from moving toward the contact portion between the transfer film and the separation portion through the bottleneck formed by the electrode cover and the first recess. Can do.

In the biomolecule analyzer according to Aspect 6 of the present invention, in any one of Aspects 1 to 5,
Said 1st recessed part is provided in the lower part of the edge part by the side of the said electrode in the said contact location.

  According to said structure, it is prevented that the electric force line produced from the contact location in a separation part at the time of electrophoresis spreads to the electrode side. As a result, the separation performance of the specimen can be increased.

In the biomolecule analyzer according to Aspect 7 of the present invention, in any one of Aspects 1 to 6,
The surface facing the buffer solution tank in the electrode cover is inclined with respect to the transport direction,
An end farther from the contact location on the surface facing the buffer solution tank is away from the electrode in a direction perpendicular to the in-plane direction of the transfer film than an end closer to the contact location on the electrode cover. It is characterized by being farther away.

  According to said structure, the bubble generate | occur | produced from the electrode can be escaped in the direction away from a contact location.

In the biomolecule analyzer according to Aspect 8 of the present invention, in any one of Aspects 1 to 7,
The electrode cover has a side wall portion that extends toward the bottom surface of the buffer solution tank along two sides parallel to the transport direction.

  According to the above configuration, it is possible to prevent the bubbles generated from the electrodes from moving from the two sides along the transfer film transport direction in the electrode cover toward the contact portion between the transfer film and the separation portion. it can.

In the biomolecule analyzer according to Aspect 9 of the present invention, in any one of Aspects 1 to 8,
The water contact angle on the surface of the electrode cover is 90 ° or less.

  According to said structure, the bubble generate | occur | produced from the electrode can be made easier to escape along the surface of an electrode cover.

  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 suitably used for a two-dimensional electrophoresis apparatus.

1 Transfer membrane 30 Anode buffer tank (buffer tank)
30a, 30a '1st recessed part 30b 2nd recessed part 32 Anode (electrode)
35, 36, 37, 38 Anode cover (electrode cover)
35a End side (end)
36a, 37a, 37a ', 38a, 38a' Convex part 41 Cathode 50 Electrophoresis gel chip (separation part)
50a Discharge unit 52 Separation gel 100, 101, 102, 103, 104 Biomolecule analyzer

Claims (9)

A buffer bath;
A transfer film disposed in the buffer solution tank;
A transport unit that transports the transfer film along a predetermined transport direction;
A separation unit that vertically contacts the transfer film and is vertically installed, separates the specimen by electrophoresis, and discharges the separated specimen to the transfer film;
An electrode disposed at a position away from the contact portion between the separation part and the transfer film in the transport direction by a certain distance;
An insulating electrode cover disposed between the electrode and the contact portion;
A first recess is provided at a position sandwiched between the electrode and the contact portion at the bottom of the buffer tank,
The biomolecule analyzer characterized in that the electrode cover hangs down toward the first recess. The buffer tank is provided with a second recess at a position away from the first recess by a certain distance in the transport direction,
The biomolecule analyzer according to claim 1, wherein the electrode is disposed in the second recess.   The biomolecule analyzer according to claim 1, wherein the electrode cover includes a convex portion that hangs down toward the first concave portion. The convex portion in the electrode cover is composed of a plurality of convex portions,
The first recess in the buffer tank is composed of a plurality of recesses,
4. The biomolecule analyzer according to claim 3, wherein the plurality of convex portions are arranged so as to individually hang down from the plurality of concave portions.   The biomolecule analyzer according to any one of claims 1 to 4, wherein an end of the electrode cover or the convex part is loosely inserted into the first concave part. .   The biomolecule analyzer according to any one of claims 1 to 5, wherein the first recess is provided at a lower portion of the end portion on the electrode side in the contact portion. The surface facing the buffer solution tank in the electrode cover is inclined with respect to the transport direction,
An end farther from the contact location on the surface facing the buffer solution tank is away from the electrode in a direction perpendicular to the in-plane direction of the transfer film than an end closer to the contact location on the electrode cover. The biomolecule analyzer according to any one of claims 1 to 6, wherein the biomolecule analyzer is further away.   The said electrode cover has a side wall part which spreads toward the bottom face of the said buffer solution tank along two sides parallel to the said conveyance direction, The any one of Claims 1-7 characterized by the above-mentioned. Biomolecule analyzer.   The biomolecule analyzer according to any one of claims 1 to 8, wherein a contact angle with water on the surface of the electrode cover is 90 ° or less.

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