TW201321398A - Sugar chain refinement device and sugar chain refinement method - Google Patents

Abstract

A sugar chain purifying device and a sugar chain purifying method each capable of easily separating and purifying many sugar chains at an excellent accuracy are provided. This sugar chain purifying device 1 is a device for purifying sugar chains from a sugar chain containing liquid. The sugar chain purifying device 1 includes: a plate assembly 10 composed of a first upper plate 100, a second upper plate 100', a first lower plate 300, a second lower plate 200 and a support body 700 for supporting them in a state that one of the first upper plate 100 and the second upper plate 100' and one of the first lower plate 300 and the second lower plate 200 are arranged at upper and lower sides of the support body 700 and assembled thereto; and a stage 3 on which the plate assembly 10 is placed. According to such a plate assembly 10, on the stage 3, the first upper plate 100 and the second upper plate 100' are changed to each other and the first lower plate 300 are the second lower plate 200 are changed to each other.

Description

Sugar chain purification device and sugar chain purification method

The present invention relates to an apparatus and method for purifying a sugar chain possessed by a glycoprotein.

The sugar chain is a monosaccharide such as glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine, sialic acid, and the like. A generic term for a molecule that is glycosidically bonded and complexed into a chain.

Sugar chains are extremely diverse and are substances associated with the various functions of naturally occurring organisms. The sugar chain is present in the form of a complex saccharide bound to a protein or a lipid, and is one of important constituents in the living body. The industry is gradually becoming more aware of the deep correlation between sugar chains in organisms and cell-to-cell signaling, protein function or interaction adjustment.

Examples of the biopolymer having a sugar chain include a proteoglycan which is a cell wall of a plant cell which contributes to stabilization of cells, and affects cell differentiation, cell proliferation, cell adhesion, cell movement, and the like. Glycolipid, glycoprotein associated with cell-cell interaction or cell recognition, and the like. The industry is gradually clearing the mechanism by which these polymeric sugar chains can control the precise biological reaction while performing, assisting, amplifying, regulating, or inhibiting functions. Further, if the relationship between the sugar chain and the differentiation and proliferation of cells, cell adhesion, immunity, and canceration of cells is clearly defined, it is expected that the sugar chain engineering is closely related to medicine, cell engineering, or organ engineering. And seeking new development (Non-Patent Document 1).

It is known that sugar chains present on the cell surface in particular play an important role as a basis for the reaction of various organisms. For example, it is said that a sugar chain is involved in the occurrence of a disease caused by an abnormal interaction with a receptor, or an infection such as an HIV or influenza virus, a toxin of a pathogenic coliform O 157 or a cholera toxin into a cell. Further, in certain cancer cells, a specific sugar chain appears on the cell surface. Thus, it is considered that the sugar chain on the cell surface imparts an important molecule to the cell.

In order to analyze the occurrence of such diseases, the industry is developing a sugar chain structure analysis technology. These techniques combine the steps of separating the sugar chain, the separation and purification of the sugar chain, and the labeling of the sugar chain from the complex saccharide. However, these steps are extremely cumbersome.

Further, as the separation and purification of the sugar chain, for example, a method using an ion exchange resin, a method using reverse phase chromatography, a method using activated carbon, or a gel filtration chromatography method can be employed. However, these separation methods are not methods for specifically identifying sugars, and thus inclusions (peptides, proteins, etc.) are inevitably mixed. Further, depending on the structure of the sugar chain, the recovery rate of the sugar chain is often different.

Further, in the case where the sugar chain is separated with high precision by chromatography, it is necessary to carry out a fluorescent label such as pyridylation on the sugar chain, which requires an cumbersome operation. Further, when analyzing the fluorescently labeled sugar chain, the unreacted 2-aminopyridine or the like is removed from the labeled reaction solution, and the labeled sugar chain is purified.

In general, gel filtration is performed using the difference in molecular weight between the labeled sugar chain and the inclusions to remove inclusions. However, this method requires a large number of instruments and requires a lot of time for operation, which is difficult for such a short period of time. A large number of samples were processed in between.

Further, as a simple method, a method of distilling off impurities by azeotropy has been attempted, but it is difficult to sufficiently remove inclusions.

Furthermore, in order to investigate the relationship between the structure of the sugar chain and various diseases, it is necessary to study the sugar chain structure of a large number of samples so that the statistical processing of the relationship can be performed.

In the previous method as described above, in order to separate the sugar chain, it takes a complicated step, which requires a huge cost and time. Therefore, the industry has sought a method for separating and purifying a large number of sugar chains with simplicity and precision.

Further, various methods have been developed for performing a fluorescent label on a sugar chain (for example, Patent Documents 1, 2, and 3). However, the labeling efficiency was not 100%, and the labeled sugar chain and the unlabeled sugar chain were mixed in one sample. This situation is not a big problem when fluorescent detection of a sugar chain by high performance liquid chromatography (HPLC, High Performance Liquid Chromatography) or capillary electrophoresis (CE (Capillary Electrophoresis) method). However, when analyzing a sugar chain by mass spectrometry, there is a problem that a peak of a mass spectrometry or a mass chromatogram is complicated. Further, in the case where the labeling efficiency of the sugar chain is inferior, even if it is analyzed by HPLC or CE, there is a possibility that the sensitivity of detecting the sugar chain is lowered.

[Previous Technical Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. Hei 7-20131

[Patent Document 2] Japanese Patent Laid-Open Publication No. 2006-098367

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2008-309501

[Non-patent literature]

[Non-Patent Document 1] Introduction to Sugar Chain Biology Chemical Fellows Issued on November 1, 2005 First Edition

An object of the present invention is to provide a sugar chain purification device capable of separating and purifying a large number of sugar chains with ease and precision, and a sugar chain purification method using the same.

The above object is achieved by the present invention of the following (1) to (25).

(1) A sugar chain purification device comprising: a plate assembly used for purifying the sugar chain from a solution containing a sugar chain; a stage on which the plate assembly is placed; a dispensing mechanism for the nozzle that ejects the plate assembly on the stage, and a moving mechanism that moves the plate assembly on the stage relative to the nozzle, and the plate assembly includes: a first upper side plate a plate-like shape having a plurality of first holes formed by a first recess opened on the upper surface and having a first through hole penetrating the bottom of the first recess, and the above-mentioned respective first holes are covered The first through hole is provided in the first filter of the bottom portion of the first recess; the second upper plate has a plate shape, and has a second recess opened by the upper surface and is formed to penetrate the second recess. a plurality of second holes of the second through hole at the bottom and a second hole that covers the second through hole, respectively, and are provided at the bottom of the second recess and different from the first filter 2 filter; a first lower side plate having a plate shape and having one first storage portion formed by a third concave portion having an open upper surface; and a second lower side plate having a plate shape and having an open upper surface a plurality of second storage portions formed by the fourth recess; and one support body for one of the first upper side plate and the second upper side plate, the first lower side plate, and the second One of the lower side plates is placed in a state of being assembled up and down, and the plate assembly system exchanges the first upper side plate and the second upper side plate with respect to the one support body on the stage. At the same time, the first lower side plate is exchanged with the second lower side plate.

(2) The sugar chain purification device according to (1) above, further comprising a heating mechanism that heats the plate assembly.

(3) The sugar chain purification device according to (2) above, wherein the operation of the heating means is stopped when the first lower side plate and the second lower side plate are exchanged.

(4) The sugar chain purification device according to the above (2), wherein the plate assembly is in a first state in which the first upper side plate is supported by the support body and supports the first lower side plate; In the second state, the first lower side plate is exchanged with the second lower side plate in the first state, and the second lower side plate is supported by the support body; and the third state is in the second state The first upper side plate is exchanged with the second upper side plate, and the second upper side plate is supported by the support body. And the second lower side plate is exchanged with the first lower side plate to support the first lower side plate by the support body; and the fourth state is the first lower side plate and the first state in the third state 2 The lower side plates are exchanged, and the second lower side plates are supported by the support body.

(5) The sugar chain purification device according to the above (4), wherein, in the first state, each of the first holes is connected to the first storage portion via the first through hole; In the state of the first through hole, each of the first through holes communicates with each of the second storage portions; and in the third state, each of the second holes passes through the second through hole and the first through hole The storage portion is connected to each other. In the fourth state, each of the second holes communicates with each of the second storage portions via the second through holes.

(6) The sugar chain purification device according to the above (5), wherein the sugar chain purification device is configured to sequentially perform the following steps: the first step of supplying the above-mentioned respective first holes to the above a capture carrier for specifically binding a sugar chain; in a second step, the solution is separately dispensed from the nozzle into each of the first holes, and the solution is contacted with the capture carrier to capture the sugar chain to the capture a third step of removing the substance other than the sugar chain bound to the capture carrier from the pore; and the fourth step of re-freeing the sugar chain bound to the capture carrier; In a fifth step, the re-free saccharide chain is separated from the capture carrier and purified; and in the first step, the second step, the third step, and the fourth step, the plate assembly is provided In the first state, in the fifth step, the panel assembly is sequentially set to the second state, the third state, and the fourth state.

(7) The sugar chain purification device according to (6) above, wherein the heating means is changeable in each of the first step, the second step, the third step, the fourth step, and the fifth step The heating temperature of the plate assembly or the first upper plate is configured.

(8) The sugar chain purification device according to (2) above, wherein the heating means has a heater embedded in the stage.

(9) The sugar chain purification device according to (2) above, wherein the heating means is a thermostatic bath having a heater.

(10) The sugar chain purification device according to the above (1), wherein the support body has an upper support member that supports the one of the first upper side plate and the second upper side plate, and is configured separately from the upper support member. Supporting the lower support member of the one of the first lower side plate and the second lower side plate, wherein the upper support member and the lower support member support the first upper side plate and the upper side support member One of the second upper side plates is configured to be detachable/assembled in a state in which the one of the first lower side plate and the second lower side plate is supported by the lower side support member.

(11) The sugar chain purification device according to (10), wherein the exchange of the first upper side plate and the second upper side plate and the exchange of the first lower side plate and the second lower side plate respectively support the upper side support The member is removed in a state in which the lower support member is disassembled.

(12) The sugar chain purification device according to the above (1), wherein the lower surface of the one of the first upper side plate and the second upper side plate and the first lower side plate are The upper surface of the one of the second lower side plates is disposed apart from each other with a pitch of 0.5 mm or more and 10 mm or less.

(13) The sugar chain purification device according to (1) above, wherein the first filter is made of a porous film or a non-woven fabric.

(14) The sugar chain purification device according to (13) above, wherein the mesh of the first filter is 0.1 to 50 μm.

(15) The sugar chain purification device according to (1) above, wherein the second filter is made of silicone.

(16) The sugar chain purification device according to (1) above, wherein the dispensing mechanism has a pump and a tube that connects the pump to the nozzle.

(17) The sugar chain purification device according to (1) above, wherein the moving mechanism has a horizontal direction moving mechanism that moves the nozzle in a horizontal direction and a vertical direction moving mechanism that moves the nozzle in a vertical direction.

(18) The sugar chain purification device according to the above (1), wherein the first upper side plate mounting portion on which the plurality of the first upper side plates are stacked and placed on the stage is provided, and the plurality of the second upper surface is provided a second upper side plate mounting portion on which the side plates are stacked and placed, a first lower side plate mounting portion on which the plurality of first lower side plates are stacked, and a plurality of the second lower side plates are stacked and placed 2 lower side onboard Set up.

(19) The sugar chain purification device according to (1) above, further comprising a cleaning tank for cleaning the nozzle.

(20) A method for purifying a sugar chain, which comprises the step of purifying the sugar chain using a sugar chain purification apparatus according to (6) above, and in the fourth step, binding the above to a capture carrier The sugar chain is then freed and the sugar chain is labeled with a labeling reagent.

(21) The method for purifying a sugar chain according to the above (20), wherein in the fourth step, after the sugar chain is re-free from the capture carrier, the sugar chain is labeled with the labeling reagent.

(22) The method for purifying a sugar chain according to (21) above, wherein the labeling reagent is a fluorescent substance composed of an aromatic amine.

(23) The method for purifying a sugar chain according to the above (22), wherein a concentration of the fluorescent substance in the solution containing the sugar chain re-freed is 0.5 mol/L or more.

(24) The method for purifying a sugar chain according to (22) above, wherein the fluorescent substance is selected from the group consisting of 2-aminobenzamide, 2-aminobenzoic acid, 8-aminoindole-1,3,6- Trisulfonate, 8-aminonaphthalene-1,3,6-trisulphonate, 2-amino-9(10H)-acridone, 5-amino fluorescein, dansyl ethanediamine At least 1 of 7-amino-4-methylcoumarin, 3-aminobenzoic acid, 7-amino-1-naphthol, 3-(ethylamido)-6-amino acridine Kind.

(25) The method for purifying a sugar chain according to (20) above, wherein the capture carrier is a polymer particle having a mercapto group or an oxyamine group.

According to the present invention, the sugar chain can be separated and purified with ease and precision, and the sugar chain can be fluorescently labeled with good yield. Further, since the liquid (solution) supplied to the plurality of first holes or the plurality of second holes can be separately processed, a large number of sugar chains can be purified and labeled by one treatment.

Hereinafter, the sugar chain purification device and the sugar chain purification method of the present invention will be described in detail based on preferred embodiments shown in the accompanying drawings.

<First embodiment>

Fig. 1 is a perspective view showing a first embodiment of the sugar chain purification device of the present invention, Fig. 2 is a perspective view showing a plate assembly provided in the sugar chain purification device shown in Fig. 1, and Fig. 3 is a plate shown in Fig. 2. FIG. 4 to FIG. 7 are cross-sectional views taken along line BB of FIG. 2 (FIG. 4 shows a first state of the plate assembly, FIG. 5 shows a second state of the plate assembly, and FIG. 6 shows a plate assembly. In the third state, FIG. 7 shows the fourth state of the plate assembly, and FIGS. 8 to 21 are sectional views sequentially showing the steps of purifying the sugar chain by the sugar chain purification device shown in FIG. 1 (from FIG. 1 The obtained cross-sectional view is observed in the direction of the arrow A). In the following, for convenience of explanation, the upper side in FIGS. 1 to 21 will be referred to as "upper" or "upper", and the lower side will be referred to as "lower" or "lower". Further, the horizontal direction is the horizontal direction of the device (the front-rear direction) is referred to as the x-axis direction, and the horizontal direction and the direction perpendicular to the x-axis direction (the left-right direction of the device) are referred to as the y-axis direction, and the vertical direction is referred to. (Upper and lower direction) is referred to as the z-axis direction.

The sugar chain purification device (hereinafter simply referred to as "purification device") shown in Fig. 1 is used in a method for purifying a sugar chain from a sugar chain-containing solution (hereinafter referred to as "sugar chain-containing liquid") 23 to purify a sugar chain. Device. The purification device 1 is provided with a stage 3, The plate assembly 10 placed on the stage 3, the dispensing mechanism 4 having the function of dispensing liquid to the plate assembly 10 on the stage 3, and the dispensing assembly with respect to the plate assembly 10 on the stage 3 The moving mechanism 5 of the nozzle (pipette) 41 of the mechanism 4, the heating mechanism 6 for heating the plate assembly 10 on the stage 3 together with the stage 3, the cleaning tank 7 of the cleaning nozzle 41, and the dispensing nozzle The control panel 11 that controls the function of the mechanism 4, the moving mechanism 5, and the heating mechanism 6. The structure of each part will be described below.

As shown in FIGS. 2 and 3, the panel assembly 10 includes a first upper side plate (porous filter plate) 100, a second upper side plate (cleaning plate) 100', a first lower side plate (waste liquid tray) 300, and a second A lower side plate (microplate) 200, an adjustment plate (spacer plate) 600, and a support body 700. That is, the panel assembly 10 includes a plurality of first upper side plates 100, a plurality of second upper side plates 100', a plurality of first lower side plates 300, and a plurality of second lower side plates 200 shown in the stage of FIG. In the board assembly 10, the members are detachable and assembled to each other. Further, the board assembly 10 can be assembled in four types depending on the selection of each board. The four assembled states include a first state shown in FIG. 4, a second state shown in FIG. 5, a third state shown in FIG. 6, and a fourth state shown in FIG. In addition, FIG. 2 shows a set of plate assemblies 10 (hereinafter also simply referred to as "board assembly 10") placed on the board assembly mounting portion 31 of the stage 3.

In the first state, the first upper side plate 100 of the first upper side plate 100 and the second upper side plate 100' is selected, and the first lower side plate 300 of the first lower side plate 300 and the second lower side plate 200 is selected. Further, the selected first upper side plate 100 and the first lower side plate 300 are assembled so as to be placed on the upper and lower sides, and are supported by the support body 700.

In the second state, the first upper side plate 100 of the first upper side plate 100 and the second upper side plate 100' is selected, and the second lower side plate 200 of the first lower side plate 300 and the second lower side plate 200 is selected. Further, the selected first upper side plate 100 and second lower side plate 200 are assembled so as to be placed on the upper and lower sides, and are supported by the support body 700.

In the third state, the second upper side plate 100' of the first upper side plate 100 and the second upper side plate 100' is selected, and the first lower side plate 300 of the first lower side plate 300 and the second lower side plate 200 is selected. Further, the selected second upper side plate 100' and the first lower side plate 300 are assembled so as to be placed on the upper and lower sides, and are supported by the support body 700.

In the fourth state, the second upper side plate 100' of the first upper side plate 100 and the second upper side plate 100' is selected, and the second lower side plate 200 of the first lower side plate 300 and the second lower side plate 200 is selected. Further, the selected second upper side plate 100' and the second lower side plate 200 are assembled so as to be placed on the upper and lower sides, and are supported by the support body 700.

The stage 3 is a flat plate having a long length in the y-axis direction of FIG. 1, and its upper surface is divided into a plurality of regions. As shown in FIG. 1 , the stage 3 is divided into a plate assembly mounting portion 31 , a first upper side plate mounting portion 32 , a second upper side plate mounting portion 33 , a first lower side plate mounting portion 34 , and a second lower side plate. The mounting portion 35, the cleaning tank mounting portion 36, and the processing liquid mounting portion 37.

The board assembly mounting portion 31 is a region that is placed at a substantially central portion in the longitudinal direction of the stage 3 and on which one set of the board assembly 10 is placed. The sugar chain is purified from the sugar chain-containing liquid 23 contained in the plate assembly 10 placed on the plate assembly mounting portion 31.

The first upper side plate mounting portion 32 is located on the stage 3 toward the purification device 1 of FIG. The area on the right side of the front side and the one or more upper upper side plates 100 are stacked. Each of the first upper side plates 100 placed on the first upper side plate mounting portion 32 is not used by the user for exchange.

The second upper side plate mounting portion 33 is a region that is placed on the right side of the purification device 1 of FIG. 1 and is placed on the right side of the first upper side plate mounting portion 32 and overlaps the one or more second upper side plates 100'. Each of the second upper side plates 100' placed on the second upper side plate mounting portion 33 is not used by the user for exchange.

The first lower side panel mounting portion 34 is a region that is placed on the lower side of the purification device 1 and the second upper side plate mounting portion 33 of FIG. 1 and that is stacked on the first lower side plate 300. Each of the first lower side plates 300 placed on the first lower side plate mounting portion 34 is not used by the user for exchange.

The second lower side plate mounting portion 35 is a region that is placed on the lower side of the purification device 1 of FIG. 1 and the first upper side plate mounting portion 32, and is placed on the second lower side plate 200. Each of the second lower side plates 200 placed on the second lower side plate mounting portion 35 is not used by the user for exchange.

Further, in the purification apparatus 1, the exchange of the respective sheets is performed by manual operation of an operator who operates the purification apparatus 1.

Further, the detailed configuration of the board assembly 10 will be described below.

The cleaning tank mounting portion 36 is located in the region on the left side of the stage 3 and on which the cleaning tank 7 is placed, toward the purification apparatus 1 of Fig. 1 . The cleaning tank 7 placed on the cleaning tank mounting portion 36 is fixed by a fixing mechanism not shown.

The treatment liquid mounting unit 37 is located on the left side of the purification apparatus 1 and the cleaning tank mounting unit 36 of FIG. 1 and is placed on the perforated plate 81 and the storage tanks 82 to 87 in which various liquids (treatment liquids) are stored. region.

The constituent material of the stage 3 is not particularly limited, and for example, various metal materials such as stainless steel, aluminum, or aluminum alloy can be used.

The dispensing mechanism 4 has a nozzle 41, a nozzle head 44 that supports/fixes the nozzle 41, a pump 42, and a tube 43 that connects the nozzle 41 and the pump 42.

The nozzle 41 is constituted by a tubular member whose front end (lower end) is open. The nozzle 41 can suck or discharge liquid through the front end opening portion 411 by the operation of the pump 42. In the configuration shown in FIG. 1, the number of the nozzles 41 is one. However, the number of the nozzles 41 is not limited thereto. For example, a plurality of nozzles may be disposed.

The pump 42 is, for example, a gear pump or an impeller pump, and is disposed on the back side of the wall portion 12 that is erected from the stage 3 .

The tube 43 connects the nozzle 41 to the pump 42. Thereby, the nozzle 41 and the pump 42 communicate via the tube 43. Further, the tube 43 has flexibility, and when the nozzle 41 is moved by the operation of the moving mechanism 5, the movement can be followed.

The moving mechanism 5 has an x-axis direction moving mechanism (horizontal direction moving mechanism) 51, a y-axis direction moving mechanism (horizontal direction moving mechanism) 52, and a z-axis direction moving mechanism (vertical direction moving mechanism) 53. The x-axis direction moving mechanism 51, the y-axis direction moving mechanism 52, and the z-axis direction moving mechanism 53 are respectively, for example, a motor (not shown), a ball screw coupled to the motor (not shown), and a ball screw. Linked linear guides (not shown).

Further, the nozzle 41 is supported by the z-axis direction moving mechanism 53 via the nozzle head 44, and is movable in the z-axis direction by the z-axis direction moving mechanism 53.

Further, the x-axis direction moving mechanism 51 collectively supports the z-axis direction moving mechanism 53 and the nozzle 41. Thereby, the nozzle 41 can be along with the z-axis direction moving mechanism 53 along the x-axis Move in direction.

Further, the y-axis direction moving mechanism 52 collectively supports the z-axis direction moving mechanism and the nozzle 41 via the x-axis direction moving mechanism 51. Thereby, the nozzle 41 can be moved in the y-axis direction together with the x-axis direction moving mechanism 51 and the z-axis direction moving mechanism 53.

According to the moving mechanism 5 having the above configuration, the nozzle 41 can be reciprocated between the plate assembly 10 and the perforated plate 81 and the accumulators 82 to 87.

The heating mechanism 6 has a heater 61 that is embedded directly under the board assembly mounting portion 31 of the stage 3 . The heater 61 is formed by winding a heating wire such as a nichrome wire which is heated by energization into a ring shape. Further, the area of the heater 61 is larger than the area of the panel assembly 10 in plan view. That is, the panel assembly 10 is disposed inside the heater 61 in plan view.

When the heater 61 is operated and heat is generated, the heat is sequentially applied to the stage 3 and the support body 700 of the board assembly 10 (second support stage (lower side support member) 500, first support stage (upper side support) Component) 400) Transfer. Thereby, the liquid on the first upper side plate 100 can be heated to evaporate the liquid (see FIGS. 9(5) and 12(14)).

Further, by embedding the heater 61 in the stage 3, it is possible to reliably prevent the operator of the purification apparatus 1 from accidentally touching the heater 61. Thereby, even if the heater 61 is heated, it is possible to prevent or suppress the operator from being burnt.

Further, the heating mechanism 6 detects the temperature on the board assembly placing portion 31 of the stage 3. Further, based on the detection result, the voltage applied to the heater 61 can be controlled. Thereby, in each of the steps (the first step, the second step, the third step, the fourth step, and the fifth step) performed by the purification device 1, the variable Further, the heating temperature of the panel assembly 10 is set to an optimum temperature, which contributes to purification of the sugar chain with excellent precision.

The cleaning tank 7 is composed of a member having a bottomed cylindrical shape, and the cleaning liquid 71 of the cleaning nozzle 41 can be stored inside. Each of the nozzles 41 is inserted into the cleaning tank 7 from the plate assembly 10 to the perforated plate 81 or the storage tanks 82 to 87, and is cleaned by the cleaning liquid 71 in the cleaning tank 7. Thereby, for example, contamination on the board assembly 10 can be prevented.

Further, in the purification apparatus 1, the cleaning liquid 71 is replaced with a non-user every time the nozzle 41 is cleaned, that is, every time it is used. Thereby, the above contamination can be surely prevented.

The cleaning liquid 71 is not particularly limited, and for example, purified water (distilled water) or an alcohol represented by methanol or ethanol can be used. Further, two or more cleaning tanks 7 may be provided on the stage 3, and the cleaning liquids 71 may be continuously washed by two or more cleaning liquids 71 stored in the two or more cleaning tanks 7.

The control panel 11 is disposed in the vicinity of the first upper side plate mounting portion 32 on the stage 3. A liquid crystal screen 111 as a display unit and an operation unit (input unit) is provided on the control panel 11. Further, for example, an input screen for setting various operating conditions of the purification device 1 or the like is displayed on the liquid crystal screen 111. The operator can set various operating conditions by touching the liquid crystal screen 111 with a finger. With this setting, the operations of the dispensing mechanism 4, the moving mechanism 5, and the heating mechanism 6 are controlled.

As described above, one set of the plate assembly 10 is placed on the board assembly mounting portion 31 of the stage 3. As shown in FIGS. 2 and 3, the panel assembly 10 includes a first upper side plate 100, a second upper side plate 100', a first lower side plate 300, and a second lower side plate 200. The whole plate 600 and the support body 700. Further, the panel assembly 10 can be in four assembled states of the first state shown in FIG. 4, the second state shown in FIG. 5, the third state shown in FIG. 6, and the fourth state shown in FIG. Further, according to the above-described four types of assembly states, the support body 700 supports the first upper side plate 100 or the second upper side plate 100' by the first support base 400, and supports the first lower side plate 300 or the second lower side plate by the second support base 500. 200.

As shown in FIG. 3, the first upper side plate 100 has a flat shape as a whole, and includes a plurality of recesses (first recesses) which are provided in the thickness direction (vertical direction), that is, the upper surface 108 is opened (the first recess). In the present embodiment, there are 96 holes (first holes) 101. The holes 101 are arranged in a matrix (12 rows in the x-axis direction and 8 rows in the y-axis direction), and are used when purifying the sugar chain from the sugar chain-containing liquid 23.

As shown in FIGS. 3 and 4, the holes 101 have a bottom portion 103 on the lower side thereof, and a through hole (first through hole) 104 penetrating the bottom portion 103 at a substantially central portion of the bottom portion 103. Thereby, the liquid supplied (accommodated) in the hole 101 flows out from the bottom 103 to the outside of the hole 101 through the through hole 104.

The diameter of the through hole 104 is preferably set to about 0.1 to 1 mm, and more preferably set to about 0.2 to 0.7 mm.

Further, a membrane-shaped filter (first filter) 105 is disposed on the lower side (the bottom 103 side) of each of the holes 101 so as to block (cover) the through holes 104. By the relationship between the diameter of the filter 105 and the through hole 104, when the first upper side plate 100 is left still, the liquid supplied into the hole 101 is prevented from flowing out (obstructed) to the outside of the hole 101 through the through hole 104. On the other hand, as described below, the first upper side plate 100 is attached to the first support base 400, and suction is used. When the pump 13 suctions the inside of the recessed portion 505 of the second support base 500, the liquid is allowed to permeate (pass) the filter 105. Therefore, the liquid flows out from the bottom 103 side to the outside of the hole 101 through the through hole 104.

The material of the filter 105 is not particularly limited, and examples thereof include a porous film and a nonwoven fabric. In addition, as a constituent material, polytetrafluoroethylene, cellulose ester, vinylidene fluoride, polycarbonate, polyethylene, polypropylene, and nylon may be used, and one type or a combination of two or more types may be used. use.

Further, it is preferred that the filter 105 be subjected to a hydrophilic treatment. Thereby, the improvement of the permeability of the above liquid can be achieved. The hydrophilic treatment can be modified by, for example, surface modification represented by plasma treatment, corona discharge, graft treatment, acid treatment, or the like, or imparting (coating, etc.) a surfactant, a water-soluble hydrazine, and a polymerization to the filter 105. It is carried out by treatment with propylene glycol or the like.

The pore diameter of the pores of the filter 105, that is, the mesh of the filter 105 is preferably set to about 0.1 to 50 μm, more preferably about 0.2 to 20 μm, and further preferably set to 0.4 to 10 μm. . Thereby, the permeation of the liquid component contained in the liquid is allowed, and at the same time, the transmission of the polymer particles 20 as described below is surely not allowed.

The second upper side plate 100' has a flat shape as a whole, and includes a plurality of recesses (second recesses) provided in the thickness direction (vertical direction), that is, the upper surface 108' is opened (in the present embodiment) It is 96) holes (2nd holes) 101'. The holes 101' are arranged in a matrix (12 rows in the x-axis direction and 8 rows in the y-axis direction) in the same manner as the holes 101 of the first upper plate 100, and the sugar chain is purified from the sugar chain-containing liquid 23 use.

As shown in FIGS. 6 and 7, the holes 101' each have a bottom portion 103' on the lower side thereof, and further have a through hole (second through hole) 104 penetrating the bottom portion 103' at a substantially central portion of the bottom portion 103'. '. Thereby, the liquid supplied (accommodated) in the hole 101' flows out from the bottom 103' to the outside of the hole 101' via the through hole 104'.

Further, a membrane-shaped filter (second filter) 106 is disposed on the lower side (the bottom 103 side) of each of the holes 101 so as to block (cover) the through holes 104. Thereby, when the second upper side plate 100' is left, the liquid supplied into the hole 101' is prevented from flowing out (blocked) to the outside of the hole 101' via the through hole 104'. On the other hand, when the second upper side plate 100' is attached to the first support base 400 and the suction pump 13 suctions the inside of the recessed portion 505 of the second support base 500, the liquid is allowed to permeate (see below). Passing the filter 106. Therefore, the liquid flows out from the side of the bottom portion 103' to the outside of the hole 101' via the through hole 104'.

The filter 106 is composed of a constituent material different from the filter 105, and has a different capturing function for an object such as a sugar chain, that is, a capturing object. The filter 106 is composed, for example, of silicone rubber.

The first upper side plate 100 and the second upper side plate 100' as described above are attached to the first support base 400 of the support body 700.

As shown in FIG. 3 to FIG. 7 , the first support base 400 has a flat shape, and has a bottom portion 401 having an opening portion 402 having a central portion opening, and an upper outer wall 403 is provided to protrude upward along the outer edge of the bottom portion 401. And the lower side outer wall 404 is provided so as to protrude downward along the outer edge of the bottom portion 401.

Further, the concave portion 405 is formed on the first support base 400 by the upper outer wall 403. By inserting the first upper side plate 100 or the second upper side plate 100' into the recessed portion 405, the inserted first upper side plate 100 is supported by the first support base 400 or The second upper side plate 100'.

Further, a groove is provided along the edge portion of the opening portion 402 of the bottom portion 401. A gasket (sealing member) 407 is disposed corresponding to the groove. Thereby, when the first upper side plate 100 or the second upper side plate 100' is inserted into the recessed portion 405, the communication between the opening portion 402 and the outside thereof is blocked.

Further, the upper outer wall 403 has an opening 408 in a direction orthogonal to the thickness direction (vertical direction) at a central portion of the two long sides facing each other. Thereby, when the first upper side plate 100 or the second upper side plate 100' inserted (placed) in the concave portion 405 is taken out, the finger can be inserted into the opening portion 408 and inserted into the upper side plate of the concave portion 405. This is lifted up so that the take-out operation can be easily performed.

Further, the concave portion 406 is formed on the first support base 400 by the lower outer wall 404. The protruding portion 502 of the second support base 500 described below is inserted along the inner circumferential surface of the recess 406, whereby the first support base 400 is fixed to the second support base 500.

In the present embodiment, the outer peripheral surface of the upper outer wall 403 and the outer peripheral surface of the lower outer wall 404 are integrally formed, and the flat outer surface is formed by one flat surface.

As shown in FIG. 4 and FIG. 6, the first lower side plate 300 has a flat shape as a whole, and has one concave portion (third concave portion) (first storage portion) 301 whose upper surface 302 is open.

As shown in FIG. 3, the first lower side plate 300 has a concave portion forming portion 303 in which the concave portion 301 is formed, and a frame portion 304 that is disposed to surround the concave portion forming portion 303. The thickness of the concave portion forming portion 303 is thicker than the frame portion 304. Thereby, the upper surface 302 of the concave portion forming portion 303 protrudes from the upper surface of the frame portion 304. The shape of the recess forming portion 303 is set to be insertable into the first support table 400. The size within the opening 402. Therefore, in a state in which the first lower side plate 300 is disposed in the recessed portion 505 of the second support base 500 (see FIGS. 4 and 6), when the first support base 400 is placed on the second support base 500, the concave portion can be formed. The forming portion 303 is inserted into the opening portion 402 of the first support table 400.

Further, as described above, the upper surface 302 of the concave portion forming portion 303 protrudes from the upper surface of the frame portion 304. However, the present invention is not limited thereto, and may be the same height as the upper surface of the frame portion 304. In this case, the purified sugar chain can also be reliably separated.

As shown in FIG. 5 and FIG. 7 , the second lower side plate 200 has a flat shape as a whole in the same manner as the first lower side plate 300 , and includes a recess opened in the thickness direction (vertical direction) by the upper surface 204 ( A plurality of (96 in the present embodiment) holes (second storage portions) 201 formed by the fourth recesses. The holes 201 are arranged in a matrix (12 rows in the x-axis direction and 8 rows in the y-axis direction).

The holes 201 are provided with a bottom portion 203 on the lower side thereof, respectively. Thereby, the liquid supplied (accommodated) into the hole 201 is stored therein.

Further, as shown in FIG. 3, the second lower side plate 200 has a hole forming portion 202 in which a plurality of holes 201 are formed, and a frame portion 205 which is disposed to surround the periphery of the hole forming portion 202. The thickness of the hole forming portion 202 is thicker than the frame portion 205. Thereby, the upper surface of the hole forming portion 202 protrudes from the upper surface of the frame portion 205. The shape of the hole forming portion 202 in plan view is set to be sized to be inserted into the opening 402 of the first support table 400. Therefore, in a state in which the second lower side plate 200 is disposed in the recessed portion 505 of the second support base 500 (see FIGS. 5 and 7), when the first support base 400 is placed on the second support base 500, the hole can be made. The forming portion 202 is inserted into the opening portion 402 of the first support table 400.

The adjustment plate 600 is used to insert the recessed portion 505 of the second support base 500 before the first lower side plate 300 or the second lower side plate 200.

The adjustment plate 600 has a flat shape and is prepared to have a plurality of different thicknesses.

Further, by selecting the adjustment plate 600 of an appropriate thickness according to the type of the first lower side plate 300 and the second lower side plate 200 inserted in the recessed portion 505, the following effects can be obtained. In the first state, the first upper side plate 100 and the first lower side plate 300 can be brought as close as possible. In the second state, the first upper side plate 100 and the second lower side plate 200 can be brought as close as possible. In the third state, the second upper side plate 100' and the first lower side plate 300 can be brought as close as possible. In the fourth state, the second upper side plate 100' and the second lower side plate 200 can be brought as close as possible. In particular, in the second state, the first upper side plate 100 and the second lower side plate 200 can be brought as close as possible, and in the fourth state, the second upper side plate 100' and the second lower side plate 200 can be brought as close as possible to each other. The situation is valid in the following steps.

The second support stand 500 is a member for supporting the first lower side plate 300 or the second lower side plate 200. As shown in FIGS. 4 to 7, the second support base 500 is formed separately from the first support base 400, and has a flat bottom portion 501 and an outer wall 503 that protrudes upward along the outer edge of the bottom portion 501. .

By enclosing the outer wall 503, a recess 505 is formed inside the outer wall 503. The first lower side plate 300 or the second lower side plate 200 is supported by the second support base 500 by inserting the first lower side plate 300 or the second lower side plate 200 into the recessed portion 505.

Further, the outer wall 503 has a protruding portion 502 that protrudes upward along the inner edge of the outer wall 503. The protruding portion 502 is set to be inserted along the inner circumferential surface of the concave portion 406 provided in the first support base 400. Thereby, on the second support table When the first support base 400 is placed on the 500, the first support base 400 is fixed to the second support base 500 by inserting the protruding portion 502 into the recessed portion 406 of the first support base 400.

A groove is provided on the upper surface of the outer wall 503 so as to surround the protrusion 502. A gasket (sealing member) 507 is disposed corresponding to the groove. When the first support base 400 is placed on the second support base 500, the communication between the inside and the outside of the panel assembly 10 is blocked between the first support base 400 and the second support base 500.

Further, as shown in FIG. 3, the outer wall 503 has a through hole 506 penetrating the outer wall 503 on the side surface of the outer wall 503. As shown in Fig. 1, the through hole 506 is connected to the suction pump 13 via a tube 14, and the suction pump 13 is constituted by a vacuum pump, and is disposed adjacent to the pump 42 and disposed on the back of the wall portion 12 which is erected from the stage 3. side. Thereby, the inside of the recessed portion 505 can be decompressed by operating the suction pump 13.

In addition, the constituent materials of the first upper side plate 100, the second upper side plate 100', the first lower side plate 300, and the second lower side plate 200 are not particularly limited, and examples thereof include polypropylene, polyethylene, and polystyrene. A resin material such as a polyvinyl chloride or a polytetrafluoroethylene may be used alone or in combination of two or more.

Further, as the constituent material of the first support base 400 and the second support base 500, the above resin material can be used. Examples of the constituent material include Fe-based alloys such as stainless steel, Cu or Cu-based alloys, and metal-based materials of Al or Al-based alloys, such as ceramics of alumina, apatite, and aluminum nitride. One type or a combination of two or more types is used. By using a metal material as a constituent material of the first support base 400 and the second support base 500, heat from the heater 61 is sequentially transmitted to the stage 3, the second support base 500, and the first support base 400. The liquid on the first upper side plate 100 can be surely heated and evaporated (see FIGS. 9(5) and 12(14)).

Next, the first state, the second state, the third state, and the fourth state in the panel assembly 10 will be described. Which of the first state, the second state, the third state, and the fourth state is the state in which the plate assembly 10 is subjected to the steps of purifying the sugar chain by the purification device 1 described below. Choose from a variety of steps.

Further, in the panel assembly 10, the distance between the holes 101 of the first upper side plate 100, the distance between the holes 101' of the second upper side plate 100', and the distance between the holes 201 of the second lower side plate 200 are set. They are the same size as each other.

As shown in FIG. 4, in the first state, the first upper side plate 100 is inserted into the recessed portion 405 of the first support base 400, and the adjustment plate 600 and the first lower side plate 300 are inserted into the recessed portion 505 of the second support base 500. . In this state, the first support base 400 is placed on the second support base 500. At this time, the concave portion forming portion 303 of the first lower side plate 300 is inserted into the opening portion 402 of the first support base 400, and the protruding portion 502 of the second support base 500 is inserted into the concave portion 406 of the first support base 400.

By the above assembly, each of the holes 101 of the first upper side plate 100 communicates with the concave portion 301 of the first lower side plate 300 via the through hole 104. In this state, the first support base 400 is fixed to the second support base 500.

Further, in this state, the communication between the inside and the outside of the panel assembly 10 is blocked between the first support base 400 and the second support base 500 by the washer 507. Further, the communication between the first support base 400 and the first upper side plate 100 is blocked by the washer 407. Thereby, the internal space 9a formed (formed) by the first upper side plate 100, the first support stage 400, and the second support stage 500 can be made An enclosed space that is closed to the outside.

Further, by operating the suction pump 13 coupled to the through hole 506 to decompress the internal space 9a (the recess 505), the internal space 9a can be made negative with respect to the outside. Therefore, a pressure difference is generated in each of the holes 101 of the first upper side plate 100 via the filter 105, whereby the liquid in the hole 101 is transmitted through the filter 105 by the pressure difference. Therefore, the liquid can flow out from the respective holes 101 through the through holes 104, and the liquid can be collectively stored in the concave portion 301 of the first lower side plate 300.

Further, in the present embodiment, the lower side of the filter 105 is a negative pressure from the upper side, and the liquid in the hole 101 is transmitted through the filter 105 by the pressure difference. However, the present invention is not limited thereto, and for example, The centrifugal force or gravity imparted to the liquid located on the upper side of the filter 105 is utilized. For example, when the viscosity of the liquid in the hole 101 is relatively low and the liquid is permeable to the filter 105, the liquid can be allowed to pass through the filter 105 by gravity.

As shown in FIG. 5, in the second state, the first upper side plate 100 is inserted into the recessed portion 405 of the first support base 400, and the adjustment plate 600 and the second lower side plate 200 are inserted into the recessed portion 505 of the second support base 500. . In this state, the first support base 400 is placed on the second support base 500. At this time, the hole forming portion 202 of the second lower side plate 200 is inserted into the opening portion 402 of the first support base 400, and the protruding portion 502 of the second support base 500 is inserted into the concave portion 406 of the first support base 400.

By the above assembly, the holes 101 of the first upper side plate 100 communicate with the holes 201 of the second lower side plate 200 via the through holes 104, respectively. In this state, the first support base 400 is fixed to the second support base 500.

Further, in the second state, the internal space 9a may be a closed space that is closed to the outside. Further, by operating the suction pump 13 coupled to the through hole 506 to decompress the internal space 9a (the recess 505), it is possible to flow out from the liquid holes 101 through the through holes 104. As a result, the liquid flowing out from each of the holes 101 can be independently stored in the holes 201 of the second lower side plate 200, respectively.

As shown in FIG. 6, in the third state, the second upper side plate 100' is inserted into the recessed portion 405 of the first support base 400, and the adjustment plate 600 and the first lower side plate are inserted into the recessed portion 505 of the second support base 500. 300. In this state, the first support base 400 is placed on the second support base 500. At this time, similarly to the first state, the concave portion forming portion 303 of the first lower side plate 300 is inserted into the opening portion 402 of the first support base 400, and the protruding portion 502 of the second support base 500 is inserted into the first support table 400. Inside the recess 406.

By the above assembly, each of the holes 101' of the second upper side plate 100' communicates with the concave portion 301 of the first lower side plate 300 via the through hole 104'. In this state, the first support base 400 is fixed to the second support base 500.

Further, in this state, the communication between the inside and the outside of the panel assembly 10 is blocked between the first support base 400 and the second support base 500 by the washer 507. Further, the communication between the first support base 400 and the second upper side plate 100' is blocked by the washer 407. Thereby, the internal space 9b formed (formed) by the second upper side plate 100', the first support base 400, and the second support stand 500 can be closed to the outside.

Further, by operating the suction pump 13 coupled to the through hole 506 to decompress the internal space 9b (the recess 505), the internal space 9b can be made negative with respect to the outside. Therefore, in each hole 101' of the second upper side plate 100', respectively A pressure difference is generated above and below the filter 106, whereby the liquid in the hole 101' is transmitted through the filter 106 by the pressure difference. Therefore, the liquid can flow out from the respective holes 101' through the through holes 104', so that the liquid can be collectively stored in the concave portion 301 of the first lower side plate 300.

As shown in FIG. 7, in the fourth state, the second upper side plate 100' is inserted into the recessed portion 405 of the first support base 400, and the adjustment plate 600 and the second lower side plate are inserted into the recessed portion 505 of the second support base 500. 200. In this state, the first support base 400 is placed on the second support base 500. At this time, the hole forming portion 202 of the second lower side plate 200 is inserted into the opening portion 402 of the first support base 400, and the protruding portion 502 of the second support base 500 is inserted into the concave portion 406 of the first support base 400.

By the above assembly, the respective holes 101' of the second upper side plate 100' communicate with the holes 201 of the second lower side plate 200 via the through holes 104'. In this state, the first support base 400 is fixed to the second support base 500.

Further, in the fourth state, the internal space 9b may be a closed space that is closed to the outside. Further, by operating the suction pump 13 coupled to the through hole 506 to decompress the internal space 9b (the recess 505), it is possible to flow out from the liquid hole 101' through the through hole 104'. As a result, the liquid flowing out from each of the holes 101' can be independently stored in the holes 201 of the second lower side plate 200, respectively.

As described above, in the panel assembly 10, the upper surface 109 of the first upper side plate 100 and the upper surface 302 of the first lower side plate 300 can be brought as close as possible to each other by the adjustment plate 600 in the first state. The distance d _{1 is} set to be as small as possible. In the second state, the lower surface 109 of the first upper side plate 100 and the upper surface 204 of the second lower side plate 200 may be as close as possible, and the distance d _{2 may be} set as small as possible. In the third state, also the upper side of the second 'below surface 109' 100 and 300 on the lower surface of the first side plate 302 as close as possible, the distance D ₃ of the other set as small as possible. In the fourth state, can the upper side of the second 'below surface 109' 100 and 200 on the lower surface of the second side plate 204 as close as possible, and the distance D _4, etc. is set as small as possible.

Further, in particular, in the second state, even if the liquid passing through the through hole 104' is scattered, the liquid can be efficiently supplied (recovered) into the hole 201 of the second lower side plate 200 corresponding to each of the holes 101. Similarly, in the fourth state, even if the liquid passing through the through hole 104' is scattered, the liquid can be efficiently supplied into the hole 201 of the second lower side plate 200 corresponding to each of the holes 101'.

In addition, the pitch d ₁ to d ₄ is not particularly limited, and is preferably set to 10 mm or less, more preferably 0.5 mm or more and 10 mm or less, and further preferably set to 0.5 mm or more and 3 Below mm. By setting the pitches d ₁ to d ₄ within the range, the above effects can be exhibited more remarkably.

Further, as shown in Fig. 7 (the same applies to Fig. 6), it is preferable that the bottom surface (lower surface 109') of the second upper side plate 100' protrudes so as to surround each of the through holes 104' to form an annular projection 107. . By forming the projections 107, the liquid passing through the through holes 104' can be guided into the holes 201 of the second lower side plate 200, so that the liquid can be recovered more efficiently.

Next, the procedure for purifying the sugar chain by the purification device 1 will be described with reference to Figs. 8 to 21, and before this, the "sugar chain" and the "capture carrier specifically binding to the sugar chain" will be described.

Here, the case where the sugar chain is obtained from the glycoprotein containing a sugar chain is one. The example is explained. The sample containing the glycoprotein is not particularly limited, and examples thereof include biological samples such as whole blood, serum, plasma, urine, saliva, cells, and tissues.

Then, the sample is subjected to, for example, SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) or an isoelectric point electrophoresis combined with SDS-PAGE. Specific processing such as two-dimensional electrophoresis. Thereby, the glycoprotein contained in the sample is separated in the gel. Thereafter, the sugar chain is freed from the glycoprotein held in the gel by a sugar chain free method. The method for releasing the sugar chain is not particularly limited, and for example, a method using glycosidase treatment using N-glycosidase or O-glycosidase, deuterium decomposition, and β detachment by alkali treatment can be employed. The gel after the sugar chain is freely treated is taken out, and the gel is washed with a washing liquid such as water. By the above treatment, the free sugar chain is eluted from the gel into the washing liquid. Then, the gel was precipitated in the washing liquid to obtain a supernatant, and the supernatant was recovered, whereby a sugar chain-containing liquid 23 was obtained. Each of the sugar chain-containing liquids 23 is stored in advance in a perforated plate 81 containing a plurality of (96 in the present embodiment) holes 811.

Further, in order to free the sugar chain, in the present embodiment, electrophoresis treatment is used, but the present invention is not limited thereto. For example, affinity chromatography treatment or ion exchange chromatography treatment may be used. Further, in the case where the operation of purifying the glycoprotein from the above sample can be omitted, the glycoprotein contained in the sample can be directly treated to release the sugar chain.

Further, the sugar chain is the only substance having an aldehyde group among the substances in the living body. That is, the sugar chain is a cyclic hemiacetal type structure and an acyclic type in an aqueous solution or the like. The aldehyde-type structure balances the substances present.

On the other hand, an organism in a living body other than a sugar chain such as a protein, a nucleic acid, or a lipid does not usually contain an aldehyde group.

Thus, it is possible to selectively capture only the sugar chain by using a capture carrier having a functional group which specifically reacts with an aldehyde group to form a stable bond.

As the functional group which specifically reacts with the aldehyde group, for example, a mercapto group, an oxyamino group, an amine group, an aminothiourea group, and the like are preferably used, and more preferably a mercapto group or an oxygen group is used. Amino group. The oxime bond formed by the reaction of the oxyamine group and the aldehyde group, and the oxime bond formed by the reaction of the thiol group and the aldehyde group can be easily cleaved by acid treatment or the like. Therefore, after the sugar chain is captured onto the capture carrier, the sugar chain can be easily cleaved from the capture vector.

Furthermore, in general, the capture/loading of physiologically active substances mostly uses an amine group, but the bond generated by the reaction of the amine group with the aldehyde group (Schiff base) is weak, so a reducing agent is required. Wait for secondary treatment. Thus, the amine group is poor for the capture of the sugar chain.

As a carrier for capturing a sugar chain, polymer particles are preferably used. Further, the polymer particles are preferably solid particles or gel particles having a functional group which specifically reacts with an aldehyde group of a sugar chain at least in one part of the surface.

When the polymer particles are solid particles or gel particles, the sugar chains are trapped on the polymer particles, and the particles can be easily recovered by using the pores 101 provided in the first upper side plate 100.

As such a polymer particle, the following formula (1) is mentioned, for example.

In the purification apparatus 1, the polymer particles 20 are used as a capture carrier that specifically binds to a sugar chain, and the sugar chain is captured onto the polymer particles 20.

The polymer particles 20 are stored in the storage tank 82 in advance in the form of the particle dispersion liquid 22 in which the polymer particles 20 are dispersed in the pure water 21 .

Further, the shape of the polymer particles 20 is not particularly limited, and for example, it is preferably spherical or a shape similar thereto. When the polymer particles 20 are spherical, the average particle diameter is preferably set to about 0.05 to 1000 μm, more preferably about 0.05 to 200 μm, and still more preferably set to about 0.1 to 200 μm. The optimum setting is about 0.1~100 μm. When the average particle diameter is less than the lower limit, it is difficult to recover the polymer particles 20 by centrifugation or filtration. Moreover, if the average particle diameter exceeds the upper limit, the polymer particles are present. The contact area between the sub- 20 and the sample solution described below is small, and the efficiency of capturing the sugar chain is lowered.

Then, in the purification apparatus 1, the following "capture carrier supply step (first step)", "sugar chain capture step (second step)", "substance removal step (third step)", and "re Free step (fourth step) and "sugar chain purification step (step 5)". Further, as needed, one or more arbitrary steps are performed between the steps.

The capture carrier supply step is a step of supplying a capture carrier (polymer particles 20) specifically bound to a sugar chain in each of the wells 101 of the first upper side plate 100. In the capture carrier supply step, the panel assembly 10 is used in the first state.

In the sugar chain capturing step, the sugar chain-containing liquid 23 is dispensed into each of the holes 101 of the first upper side plate 100 by the nozzles 41, and the sugar chain-containing liquid 23 is brought into contact with the polymer particles 20 to capture the sugar chains to the polymer particles. Steps on 20. In the sugar chain capturing step, the plate assembly 10 is continuously used in the first state after the capture carrier supply step.

The substance removing step is a step of removing substances other than the sugar chain bound to the polymer particles 20. In the substance removal step, the panel assembly 10 is used in the first state.

The re-free step is a step of re-freeing the sugar chain bound to the polymer particles 20. In the re-freeing step, the plate assembly 10 is continuously used in the first state after the substance removing step.

The sugar chain purification step is a step of separating the re-free sugar chain from the polymer particles 20 and purifying it. In the sugar chain purification step, the plate assembly 10 is sequentially changed. It is used for the second state, the third state, and the fourth state. The second state is on the stage 3, and the first lower side plate 300 and the second lower side plate 200 in the plate assembly 10 of the first state are exchanged, and the second support stand 500 (support body 700) supports the first stage. 2 lower side panel 200. In the third state, the first upper side plate 100 and the second upper side plate 100' in the second state plate assembly 10 are exchanged, and the second upper side plate 100' is supported by the first support base 400 (support body 700). The second lower side plate 200 is exchanged with the first lower side plate 300, and the first lower side plate 300 is supported by the second support stand 500 (support body 700). In the fourth state, the first lower side plate 300 and the second lower side plate 200 of the third state plate assembly 10 are exchanged, and the second lower side plate 200 is supported by the second support stand 500 (support body 700). .

[1] capture carrier supply step

[1-1 (board assembly mounting)] As shown in Fig. 8 (1), the panel assembly 10 is placed in the first state, and the panel assembly 10 is placed on the stage 3 (the board assembly is placed). Part 31). Furthermore, the operation of the heater 61 embedded in the stage 3 is still in a stopped state.

[1-2 (Pumping of Particle Dispersion)] Further, the nozzle 41 is inserted into the storage tank 82 in which the particle dispersion liquid 22 is stored, and the pump 42 is operated in this state. Thereby, the particle dispersion liquid 22 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41.

[1-3] Then, the moving mechanism 5 is operated to move the nozzle 41 sucking the particle dispersion liquid 22 onto the plate assembly 10. In addition, the particle dispersion liquid 22 (polymer particles 20) is supplied (dispensed) into the respective holes 101 of the first upper side plate 100 of the plate assembly 10 in accordance with the movement.

[2] aspiration step

Then, as shown in Fig. 8 (2), the suction pump 13 is operated to make the internal space 9a of the plate assembly 10 a negative pressure. Thereby, the particle dispersion liquid 22 in each of the holes 101 is sucked separately.

Thereby, the pure water 21 in the particle dispersion liquid 22 passes through the filter 105 in each of the holes 101. On the other hand, the polymer particles 20 cannot pass through the filter 105 due to the relationship between the pore diameter of the filter 105 and the particle diameter of the polymer particles 20. Therefore, the pure water 21 that has passed through the filter 105 is selectively supplied into the first lower side plate 300 (recessed portion 301) through the through hole 104.

As a result, as shown in FIG. 8 (3), the polymer particles 20 remain in the pores 101 of the first upper side plate 100 alone.

[3] nozzle cleaning step

Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

[4] Sugar chain capture step

[4-1 (suction of sugar chain-containing liquid)] Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the hole 811 of the perforated plate 81 in which the sugar chain-containing liquid 23 is stored, and the pump 42 is placed in this state. Operation. Thereby, the sugar chain-containing liquid 23 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41. The sugar chain-containing liquid 23 may also contain a volatile organic solvent represented by acetonitrile and an acid (acetic acid or the like) as a pH adjuster.

[4-2 (Supply of Sugar Chain-Containing Liquid)] Then, as shown in Fig. 9 (4), the moving mechanism 5 is operated to move the nozzle 41 sucking the sugar chain-containing liquid 23 to the plate group On the body 10. In addition, the sugar chain-containing liquid 23 is supplied (dispensed) to each of the holes 101 of the first upper side plate 100 of the panel assembly 10 in accordance with the movement.

[4-3 (heating of sugar chain-containing liquid)] Then, as shown in Fig. 9 (5), the heater 61 is operated. Thereby, the heat of the heater 61 is sequentially transmitted to the stage 3, the second support stage 500, the first support stage 400, and the first upper side plate 100. Thereby, the sugar chain-containing liquid 23 on the first upper side plate 100 is heated. This heating is performed to maintain the sugar chain-containing liquid 23 in a fixed temperature range (heating temperature) until the sugar chain-containing liquid 23 is dried.

Thereby, the polymer particles 20 react with the sugar chain contained in the sugar chain-containing liquid 23 to capture the sugar chain onto the polymer particles 20.

The pH of the reaction liquid (sugar chain-containing liquid 23) at this time is preferably 2 to 9, more preferably 2 to 7, and still more preferably 2 to 6. Further, the pH adjustment can be performed, for example, by adding various buffers or organic solvents to the sugar chain-containing liquid 23 in each of the pores 101 of the first upper side plate 100.

The temperature of the reaction liquid at the time of capturing the sugar chain is preferably maintained at about 4 to 100 ° C, more preferably at about 25 to 90 ° C, more preferably at about 30 to 80 ° C, and most preferably maintained at about Set the temperature range from 60 to 80 °C.

Further, when the temperature is set to the above temperature range, the reaction time, that is, the time until the sugar chain-containing liquid 23 is dried, is usually set to about 0.1 to 3 hours, preferably about 0.6 to 2 hours.

By reacting the polymer particles 20 with the sugar chain under these conditions, the sugar chain is surely captured onto the polymer particles 20.

Furthermore, as in the present embodiment, the sugar chain-containing liquid 23 is heated until sugar The chain-containing liquid 23 is dried, and the reaction rate of the polymer particles 20 and the sugar chain can be surely improved.

Further, when the polymer particles 20 are those having a mercapto group, a reaction represented by the following formula (2) is carried out between the mercapto group and the reducing end of the sugar chain, whereby the sugar chain is captured to On the polymer particles 20.

As described above, by using the panel assembly 10 of the first state, the supplied liquid can be reacted in the respective holes 101 of the first upper side plate 100. Further, by sucking the internal space 9a of the plate assembly 10, the solid component contained in the liquid supplied into the hole 101 can be easily separated from the liquid component.

[5] nozzle cleaning step

Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

[6] The first lower side board exchange step is completed

[6-1 (Disassembly of Plate Assembly)] As shown in Fig. 10 (6), the operator of the purification apparatus 1 disassembles the support body 700 of the plate assembly 10 on the stage 3 to support the first The first support base 400 of the upper side panel 100 and the second support base 500 that supports the used first (lower pure water 21) first lower side panel 300. This disassembly operation is performed by lifting the first support base 400 together with the first upper side plate 100.

[6-2 (Removal of the First Lower Side Plate)] Next, as shown in Fig. 10 (7), the used first lower side plate 300 is removed from the second support stand 500.

[6-3 (Loading of the Lower Lower Side Plate)] Next, as shown in Fig. 10 (8), the unused first lower side plate 300 is loaded into the second support base 500 of the cavity.

[6-4 (Assembly of Plate Assembly)] Next, as shown in FIG. 10 (9), the first upper side plate 100 is superposed and supported on the second support stand 500 on which the unused first lower side plate 300 is loaded. The first support base 400 again brings the board assembly 10 into the first state.

Furthermore, when this step [6] is performed, the operation of the heater 61 is stopped. Since the exchange of the first lower side panel 300 is manually operated by the operator, it is possible to surely prevent the operator from being burnt.

[7] Substance removal step

Here, when the polymer particles 20 are used as the capture carrier as described above, they are substances other than the sugar chains trapped on the polymer particles 20 (hereinafter, this substance may also be referred to as "cleaning substance"). For example, in addition to the sugar chain not captured on the polymer particles 20, inclusions (proteins, peptides, lipids, and the like) other than the sugar chain which are non-specifically adsorbed on the surface of the polymer particles 20 may be mentioned.

Therefore, in the present step [7], the substances are removed by washing with the cleaning liquid 24. The cleaning liquid 24 is not particularly limited, and examples thereof include water, various buffer solutions, various organic solvents, and the like, and these may be appropriately combined. use.

[7-1 (Suction of Cleaning Liquid)] First, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the storage tank 83 in which the cleaning liquid 24 is stored, and the pump 42 is operated in this state. Thereby, the cleaning liquid 24 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41.

[7-2 (Supply of Cleaning Liquid)] Then, as shown in Fig. 11 (10), the moving mechanism 5 is operated, and the nozzle 41 sucking the cleaning liquid 24 is moved to the plate assembly 10. Further, along with this movement, the cleaning liquid 24 is supplied (added) to each of the holes 101 of the first upper side plate 100 of the panel assembly 10.

[7-3 (Pumping of Cleaning Liquid)] Then, as shown in Fig. 11 (11), the suction pump 13 is operated to make the internal space 9a of the plate assembly 10 a negative pressure. Thereby, the cleaning liquid 24 in each of the holes 101 is sucked separately.

As a result, the cleaning liquid 24 can be passed through the filter 105. As shown in FIG. 11 (12), the cleaning liquid 24 can be selectively removed from the first upper side plate 100 to the first lower side plate 300 while the polymer particles 20 remain in the first upper side plate 100. Further, the cleaning material dissolved in the cleaning liquid 24 can also be removed.

By repeating the operations shown in FIGS. 11(10) to (12) in a plurality of times, the cleaning substance can be separated into the first lower side plate 300 while the cleaning substance is dissolved in the cleaning liquid 24. Therefore, the cleaning substance can be surely removed from the sugar chain-trapped polymer particles 20.

Further, in this case, it is preferred to first wash the polymer particles 20 sufficiently using water or a buffer as the cleaning liquid 24, and then further wash the polymer particles 20 with the cleaning liquid 24 of the organic solvent, and use the polymer particles 20 as needed. Wait for the cleaning solution 24 to be cleaned, and finally use the organic solvent cleaning solution 24 to clean the polymer. Particle 20. Thereby, the cleaning substance, in particular, the inclusions which are non-specifically adsorbed on the surface of the polymer particles 20 can be removed more surely.

[8] Nozzle cleaning step

Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

[9] Re-free step

Here, the sugar chain bound to the polymer particles 20 is released again. Further, the sugar chain is replaced with another compound (hereinafter sometimes referred to as "compound A"), that is, the sugar chain is labeled with the compound A. Further, as the compound A, a labeling reagent containing a fluorescent substance, a light absorbing material, a radioactive substance or the like is preferably used.

[9-1 (suction of compound-containing liquid)] First, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the storage tank 84 in which the compound-containing liquid 25 containing the compound A is stored, and the pump 42 is operated in this state. . Thereby, the compound-containing liquid 25 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41.

[9-2 (Supply of Compound-Containing Liquid)] Then, as shown in Fig. 12 (13), the moving mechanism 5 is operated, and the nozzle 41 sucked with the compound-containing liquid 25 is moved to the plate assembly 10. Further, along with this movement, the compound-containing liquid 25 is supplied to each of the holes 101 of the first upper side plate 100 of the panel assembly 10, respectively.

The amount of the compound A to be added to the pores 101 of the first upper side plate 100 is preferably an excess amount with respect to the polymer particles 20 having the sugar chain-trapped. Thereby, when the compound-containing liquid 25 is heated, the compound can be realized. The increase in the substitution rate of A for sugar chains.

Specifically, the amount of the compound A to be added is preferably 1.5 times or more, more preferably 3 times or more, based on the amount of the functional group which the polymer particle 20 specifically reacts with the sugar chain. Further, it is preferably set to 5 times or more, and more preferably set to 10 times or more.

[9-3 (Heating of Compound-Containing Liquid)] Next, as shown in Fig. 12 (14), the heater 61 is operated. Thereby, the compound-containing liquid 25 in the hole 101 in the first upper side plate 100 is heated. This heating maintains the compound-containing liquid 25 in a fixed temperature range (heating temperature) until the compound-containing liquid 25 is dried.

Thereby, the captured sugar chain is cleaved from the polymer particle 20, and at the same time, the compound A is added to the sugar chain. Thus, the sugar chain is labeled with Compound A. Hereinafter, the sugar chain labeled with the compound A may also be referred to as a "labeled sugar chain".

Further, the pH of the reaction liquid (the compound-containing liquid 25) at this time is preferably 2 to 9, more preferably 2 to 7, and still more preferably 2 to 6. Further, the pH adjustment can be performed by, for example, supplying the compound-containing liquid 25 to each of the holes 101 of the first upper side plate 100, and then adding various buffers to the holes 101.

The temperature of the reaction liquid at the time of labeling is preferably maintained at about 4 to 100 ° C, more preferably at about 25 to 90 ° C, still more preferably at about 30 to 80 ° C, and most preferably maintained at 60. Set the temperature range around ~80 °C.

Further, when the temperature is set to the above temperature range, the reaction time, that is, the time until the solution is dried, is usually set to about 0.1 to 3 hours, preferably. Set to 0.6~2 hours.

The sugar chain was reliably labeled with Compound A by labeling under the above conditions.

Further, as in the present embodiment, by heating the compound-containing liquid 25 until the compound-containing liquid 25 is dried, the reaction rate between the sugar chain and the compound A can be surely improved.

Further, as the compound A, a compound having an amineoxy group or a thiol group is preferably used. In the case where the polymer particles 20 are those having a mercapto group, as the compound A, it is particularly preferred to use the N-aminooxyethylene group-tryptamine group represented by the following chemical formula (3) (spermine acid A) ester).

[10] Nozzle cleaning step

Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

[11] 1st/2nd lower side panel exchange step

[11-1 (Disassembly of Plate Assembly)] As shown in Fig. 13 (15), the operator of the purification apparatus 1 disassembles the support body 700 of the plate assembly 10 on the stage 3 to support The first support base 400 of the first upper side plate 100 and the second support base 500 that supports the first lower side plate 300 in which the mixed liquid of the pure water 21 and the cleaning liquid 24 is stored. This disassembly operation is performed by lifting the first support base 400 together with the first upper side plate 100.

[11-2 (Removal of First Lower Side Plate)] Next, as shown in Fig. 13 (16), the first lower side plate 300 is removed from the second support stand 500.

[11-3 (Loading of Second Lower Side Plate)] Next, as shown in Fig. 13 (17), the second lower side plate 200 that is not used is loaded into the second support stand 500 of the cavity.

[11-4 (Assembly of Plate Assembly)] Next, as shown in FIG. 13 (18), the first upper side plate 100 is superposed and supported on the second support table 500 loaded with the unused second lower side plate 200. The first support base 400 is in the second state.

Furthermore, when this step [11] is performed, the operation of the heater 61 is stopped. Since the exchange of the first lower side plate 300 and the second lower side plate 200 is manually operated by the operator, it is possible to surely prevent the operator from being burnt.

[12] Sugar chain purification step

Here, when the labeled sugar chain is obtained by re-freezing the sugar chain captured on the polymer particle 20 as described above, the substance contained in the pore 101 of the first upper side plate 100 is used in addition to In addition to the sugar chain in which the compound A is labeled, the polymer chain 20 in which the sugar chain is free and the compound A which is not used for the labeling of the sugar chain (hereinafter sometimes referred to as "unused compound A") may be mentioned.

Therefore, in the present step [12], the purification of the labeled sugar chain is carried out by removing the polymer particles 20 and the unused compound A.

[12-1 (Pumping of Dissolving Solution)] First, the moving mechanism 5 is operated, and the nozzle 41 is inserted into a storage tank 85 in which a solution 26 which is a liquid agent capable of dissolving the dried labeled sugar chain is stored. In this state, the pump 42 is operated. Thereby, the dissolved liquid 26 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41. The solution 26 is not particularly limited, and examples thereof include water, various buffers, and various organic solvents.

[12-2 (Supply of Dissolving Solution)] Next, as shown in Fig. 14 (19), the moving mechanism 5 is operated to move the nozzle 41 sucking the dissolved liquid 26 to the plate assembly 10. Further, along with this movement, the dissolved liquid 26 is supplied to each of the holes 101 of the first upper side plate 100 of the panel assembly 10, respectively.

[12-3 (Pumping of Dissolving Solution)] Then, as shown in Fig. 14 (20), the suction pump 13 is operated to make the internal space 9a of the plate assembly 10 a negative pressure, thereby respectively sucking each The solution 26 in the well 101. Thereby, the dissolved liquid 26 passes through the filter 105 in each of the holes 101 of the first upper side plate 100.

Then, as shown in Fig. 14 (21), the polymer particles 20 remain in the respective holes 101 of the first upper side plate 100, and the solution is stored in the holes 201 of the second lower side plate 200 corresponding to the holes 101. 26. Further, at this time, the labeled sugar chain dissolved in the solution 26 may be moved into the hole 201 of the second lower side plate 200. As a result, the labeled sugar chain is separated from the polymer particles 20. In addition, since the compound A is not used, the solubility of the solution 26 is usually exhibited. Therefore, it is moved into the pores 201 of the second lower side plate 200 while being dissolved in the solution 26 .

[12-4 (Cleaning of Nozzle)] Next, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly passed through the front end opening portion 411 of the nozzle 41. The nozzle 41 is taken in and out of the nozzle 41 to clean the nozzle 41.

[12-5 (Disassembly of Plate Assembly)] As shown in Fig. 15 (22), the operator of the purification apparatus 1 disassembles the support body 700 of the plate assembly 10 on the stage 3 to support the first upper side. The first support base 400 of the side plate 100 and the second support base 500 that supports the second lower side plate 200 in which the dissolved liquid 26 is stored. This disassembly operation is performed by lifting the first support base 400 together with the first upper side plate 100.

[12-6 (Removal of the first upper side panel)] Next, as shown in Fig. 15 (23), the first support base 400 is placed beside the second support base 500, and is used from the first support base 400. The first upper side plate 100 is removed (the polymer particles 20 remain).

[12-7 (Loading of the Second Upper Side Plate)] Next, as shown in Fig. 15 (24), the second upper side plate 100' which is not used is loaded into the first support base 400 of the cavity.

[12-8 (washing liquid shifting)] Then, as shown in Figs. 16 (25) and (26), while the moving mechanism 5 is operated, the nozzles 41 are used, and the second lower side plates 200 are arranged in n rows and n rows. The solution 26 in the hole 201 is transferred to the hole 101' of the m rows and n rows (the same row as the hole 201) of the second upper side plate 100' which is not used (in the present embodiment, m is an integer of 1 to 12). , n is an integer from 1 to 8).

[12-9 (Removal of Second Lower Side Plate)] Next, as shown in Fig. 17 (27), the second lower side plate 200 is removed from the second support stand 500.

[12-10 (Loading of First Lower Side Plate)] Next, as shown in Fig. 17 (28), the unused first lower side plate 300 is loaded into the second support base 500 of the cavity.

[12-11 (Assembly of Plate Assembly)] Next, as shown in Fig. 17 (29), the second upper side plate 100' is superposed and supported on the second support stand 500 loaded with the unused first lower side plate 300. The first support base 400 is in the third state.

[12-12 (pumping of the solution)] Then, as shown in Fig. 18 (30), the suction pump 13 is operated to make the internal space 9b of the plate assembly 10 a negative pressure, thereby respectively sucking each The solution 26 in the well 101'.

At this time, since the solution 26 diluted with a nonaqueous solvent such as acetonitrile passes through the respective filters 106, the labeled sugar chain contained in the solution 26 and the unused compound A are adsorbed to the filter 106 made of silicone. The adsorption of the labeled sugar chain on the filter 106 (silicone) is higher than that of the unused compound A. Therefore, as shown in Fig. 18 (31), one portion of the unused compound A flows into (storage) together with the solution 26 into the first lower side plate 300 (recess 301). In addition, the filter 106 is in a state in which the labeled sugar chain and the unused compound A are adsorbed, and the unused compound A contained in the unused compound A-based solution 26 is not introduced into the first lower side plate 300. Use the remainder other than Compound A.

[12-13 (Pumping of Nonaqueous Solvent)] Next, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the storage tank 86 in which the nonaqueous solvent 27 such as acetonitrile is stored, and the pump 42 is operated in this state. Thereby, the nonaqueous solvent 27 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41.

[12-14 (Supply of Nonaqueous Solvent)] Next, as shown in Fig. 19 (32), the moving mechanism 5 is operated, and the nozzle 41 sucking the nonaqueous solvent 27 is moved to the plate assembly 10. Further, along with this movement, the nonaqueous solvent 27 is supplied to each of the holes 101' of the second upper side plate 100' of the panel assembly 10.

[12-15 (Pumping of Nonaqueous Solvent)] Then, as shown in Fig. 19 (33), the suction pump 13 is operated to make the internal space 9b of the plate assembly 10 a negative pressure, thereby separately sucking A non-aqueous solvent 27 in each of the holes 101'. Thereby, on the second upper side panel In each of the holes 101' of 100', the nonaqueous solvent 27 passes through the filter 106, respectively. At this time, as shown in FIG. 19 (34), the unused compound A adsorbed on the filter 106 is roughly washed and removed from the filter 106, and flows into the first lower side plate 300 together with the nonaqueous solvent 27. Further, even if a plurality of unused compounds A are adsorbed on the filter 106, the amount thereof is substantially not affected by the following measurement using the amount of the compound A by HPLC or mass spectrometry. Further, the labeled sugar chain is still in a state of being adsorbed on the filter 106.

[12-16 (Cleaning of Nozzles)] Next, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the washing tank 7 filled with the washing liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

[12-17 (Removal of Plate Assembly)] As shown in Fig. 20 (35), the operator of the purification apparatus 1 disassembles the support body 700 of the plate assembly 10 on the stage 3 to support the second The first support base 400 of the side plate 100' and the second support base 500 supporting the first lower side plate 300 that stores the mixed solution of the dissolved liquid 26 and the nonaqueous solvent 27. This disassembly operation is performed by lifting the first support base 400 together with the second upper side plate 100'.

[12-18 (Removal of First Lower Side Plate)] Next, as shown in Fig. 20 (36), the first lower side plate 300 is removed from the second support stand 500.

[12-19 (Loading of Second Lower Side Plate)] Next, as shown in Fig. 20 (37), the second lower side plate 200 that is not used is loaded into the second support stand 500 of the cavity.

[12-20 (Assembly of Plate Assembly)] Next, as shown in Fig. 20 (38), the second support table 500 loaded with the unused second lower side plate 200 is superposed and supported on the second support table 500. The first support base 400 of the second upper side plate 100' has the plate assembly 10 in the fourth state.

[12-21 (Water suction)] Then, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the storage tank 87 in which the water 28 is stored, and the pump 42 is operated in this state. Thereby, the water 28 is sucked into the nozzle 41 through the front end opening portion 411 of the nozzle 41.

[12-22 (Supply of Water)] Then, as shown in Fig. 21 (39), the moving mechanism 5 is operated, and the nozzle 41 that sucks the water 28 is moved to the plate assembly 10. Further, along with this movement, the water 28 is supplied to each of the holes 101' of the second upper side plate 100' of the panel assembly 10.

[12-23 (Water suction)] Then, as shown in Fig. 21 (40), the suction pump 13 is operated to make the internal space 9b of the plate assembly 10 a negative pressure. Thereby, the water 28 in each of the holes 101' is sucked separately. Thereby, the water 28 passes through the filter 106 in each of the holes 101' of the second upper side plate 100'. At this time, as shown in FIG. 21 (41), the labeled sugar chain in a state of being adsorbed on the filter 106 is detached from the filter 106, and flows into the hole 201 of the second lower side plate 200 together with the water 28. That is, the labeled sugar chain is purified (isolated).

[12-24 (Nozzle Cleaning)] Next, the moving mechanism 5 is operated, and the nozzle 41 is inserted into the cleaning tank 7 filled with the cleaning liquid 71, and the pump 42 is operated in this state. Thereby, the cleaning liquid 71 is repeatedly introduced into the nozzle 41 through the front end opening portion 411 of the nozzle 41, and the nozzle 41 is cleaned.

The sugar chain which was labeled with Compound A was purified by using the plate assembly 10 by the procedure as described above.

In addition, the sugar chain labeled with Compound A can be utilized as MALDI- TOF MS (Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry) is represented by mass spectrometry and further high performance liquid chromatography (HPLC). . In particular, when the sugar chain is labeled with N-aminooxyacetic acid-tryptamine (methyl arginine) represented by the above chemical formula (3), high sensitivity analysis can be carried out using MALDI-TOF MS. Further, for example, when the sugar chain is labeled with 2-aminobenzamide, high sensitivity analysis can be carried out using HPLC.

According to the above-described purification apparatus 1, the first upper side plate 100 and the second upper side plate 100' can be exchanged with respect to the plate assembly 10 on the stage 3, and the first lower side plate 300 can be exchanged with the second lower side plate 200.

In particular, as described above, the first support base 400 and the second support base 500 support the first upper side plate 100 or the second upper side plate 100' by the first support base 400, and the first support base 500 supports the first support base 500. The lower side plate 300 or the second lower side plate 200 is configured to be detachable/assembled. Further, in a state where the first support base 400 and the second support base 500 are disassembled, the first upper side plate 100 and the second upper side plate 100' can be exchanged, and the first lower side plate 300 and the second lower side can be respectively performed. The exchange of the side panels 200. Thereafter, the assembly of the assembled board 10 can also be easily performed.

In this way, the purification device 1 is configured to be easily and quickly exchanged on the stage 3 .

Thereby, the board assembly 10 on the stage 3 can be easily and surely set to any of the first state to the fourth state to be applied to each step. Therefore, a large number of sugar chains can be separated and purified simply and with high precision.

Furthermore, preferably after the [7] substance removal step is finished, or is about to enter The step of deactivating the functional groups of the capture carrier (polymer particles 20) is carried out before the step [9]. In this step, in the above step [4], a functional group (a functional group which specifically reacts with an aldehyde group of a sugar chain) which is not provided in a capture carrier for capturing a sugar chain is inactivated. Thereby, it is possible to surely prevent impurities other than the sugar chain from being chemically bonded to the capture carrier, thereby improving the purification rate of the sugar chain.

The deactivation of the functional group possessed by the capture carrier can be carried out, for example, by bringing the deactivation liquid having a function of deactivating the functional group into contact with the capture carrier and allowing it to stand. The deactivating liquid is not particularly limited, and when the functional group is a mercapto group, for example, an acid anhydride such as acetic anhydride or succinic anhydride may be mentioned. Thereby, the functional group can be easily deactivated.

The reaction conditions are preferably a deactivation liquid: 10% acetic anhydride/methanol, a temperature: normal temperature (room temperature), and a standing time: 30 minutes, but are not limited thereto. Moreover, the panel assembly 10 is used in the first state.

Further, it is preferable to perform "non-aqueous solvent supply" between [12-4 (washing of the nozzle)] and [12-5 (disassembly of the plate assembly)]. The supply of the non-aqueous solvent means that acetonitrile is supplied into each of the holes 101 of the first upper side plate 100 and acetonitrile is sucked. By the supply of the nonaqueous solvent, the labeled sugar chain remaining in the pores 101 can be recovered (rinsed) into the second lower side plate 200 without residue, and the mark stored in the second lower side plate 200 can be marked. The solution of the sugar chain was changed to an acetonitrile solution. Furthermore, by adding acetonitrile, the hydrophobicity of the solution is improved. Therefore, the labeled sugar chain (hydrophilic) is easily retained on the filter 106 (hydrophilic) composed of silicone. The labeled sugar chain is not easily retained on the filter 106 in the state of an aqueous solution. Usually, the acetonitrile content is 90% or more (preferably The acetonitrile solution was prepared in a manner of 95%).

<Second embodiment>

Fig. 22 is a perspective view showing a second embodiment of the sugar chain purification device of the present invention.

In the following, the second embodiment of the sugar chain purification device and the sugar chain purification method of the present invention will be described with reference to the drawings, and the differences from the above-described embodiments will be mainly described, and the description of the same matters will be omitted.

The configuration of the heating mechanism of the present embodiment is different, and is the same as the above-described first embodiment.

In the purification apparatus 1 of the present embodiment shown in Fig. 22, the heater 61 (heating means 6) is omitted from the stage 3. The heating mechanism 6A is replaced by a thermostatic chamber which is disposed at a position different from the stage 3 and includes a chamber 62 and a heater 63 for heating the inside of the chamber 62.

The chamber 62 has a box shape, and includes a chamber body 64 having a mouth portion 641 into which the first upper side plate 100 is inserted, and a door 65 opening and closing the opening portion 641.

Further, the heater 63 is disposed in the chamber 62, and is composed of a heating wire such as a nichrome wire which generates heat by energization.

Further, when the liquid in the hole 101 of the first upper side plate 100 is heated, the operator of the purification apparatus 1 moves the first upper side plate 100 into the chamber 62. Thereafter, the heater 63 is operated to heat the liquid.

<Third embodiment>

Fig. 23 is a perspective view showing a third embodiment of the sugar chain purification device of the present invention.

Hereinafter, the sugar chain purification device and the sugar chain purification method of the present invention will be described with reference to the accompanying drawings. The third embodiment of the method will be described, and the differences from the above-described embodiment will be mainly described, and the description of the same matters will be omitted.

The configuration of the heating mechanism of the present embodiment is different, and is the same as the above-described first embodiment.

In the purification apparatus 1 of the present embodiment shown in Fig. 23, the heater 61 (heating means 6) is omitted from the stage 3. In place of the heating mechanism 6B, a thermostatic chamber (heating block) is disposed adjacent to the plate assembly mounting portion 31 of the stage 3, and includes a heating device body 66 having a recess 661 filled with a heat medium, A heater 67 that heats the inside of the recess 661. Further, the heat medium is not particularly limited, and for example, a liquid such as water can be used.

The heating device body 66 has a recess 661 that opens toward the upper side in the drawing. The concave portion 661 may be filled with a heat medium, and the first upper side plate 100 may be housed in the heating device main body 66 while maintaining the filled state.

Further, the heater 67 is disposed on the outer side of the concave portion 661, and is composed of a heating wire such as a nichrome wire that generates heat by energization. The heat of the heater 67 is transmitted to the first upper side plate 100 via the heat medium.

Further, when the liquid in the hole 101 of the first upper side plate 100 is heated, the operator of the purification apparatus 1 moves the first upper side plate 100 into the concave portion 661 filled with the heat medium. Thereafter, the heater 67 is operated to heat the liquid.

<Fourth embodiment>

In the following, the fourth embodiment of the sugar chain purification apparatus and the sugar chain purification method of the present invention will be described, and the differences from the above-described embodiment will be mainly described, and the description of the same matters will be omitted.

In the present embodiment, in the re-freeing step, the re-freezing of the sugar chain and the labeling of the sugar chain are shifted at the time point, that is, after the sugar chain is re-free, the sugar chain is labeled, and the first one is The embodiment is the same.

In this step, as the compound A, a fluorescent substance composed of an aromatic amine is used.

In the case where the above aromatic amine is used as the compound A, the sugar chain captured on the polymer particles 20 is first separated from the polymer particles 20 and then released, and then labeled with the compound A.

Hereinafter, a case where the sugar chain is labeled with the compound A using the aromatic amine as the compound A will be described.

[9-1'] First, the sugar chain free liquid in which the sugar chain is re-free is added (supplied) to each of the holes 101 of the first upper side plate 100 containing the sugar chain-trapped polymer particles 20 by using the nozzle 41.

[9-2'] Then, the heater 61 is operated to heat the sugar chain free liquid. The sugar chain free solution is maintained at a fixed temperature range by heating the sugar chain free liquid until the added sugar chain free liquid is dried. Thereby, the captured sugar chain is cleaved from the polymer particle 20, and the sugar chain is released again.

The pH of the reaction liquid (sugar chain free solution) at this time is preferably from 2 to 9, more preferably from 2 to 7, further preferably from 2 to 6. This pH adjustment is carried out by adding a sugar chain free liquid to the pores 101 in the above step [9-1'], and various buffers are used as the sugar chain free liquid.

The temperature at which the sugar chain is free is preferably maintained at about 4 to 100 ° C, more preferably at about 25 to 90 ° C, more preferably at about 30 to 80 ° C, and most preferably at 60 to 80. Set the temperature range around °C.

Further, when the reaction time, that is, the time until the solution (sugar chain free solution) is dried, is set to the above temperature range, it is usually set to about 0.1 to 3 hours, preferably about 0.6 to 2 hours.

The sugar chain is surely cleaved from the polymer particles 20 by re-freeing the sugar chain under the above conditions.

Further, as in the present embodiment, by heating the sugar chain free liquid until the sugar chain free liquid is dried, the detachment rate of the sugar chain can be surely improved.

[9-3'] Then, using the nozzle 41, a compound-containing liquid 25 containing an aromatic amine as the compound A is added to the pores 101 of the porous filter plate 100 in which the sugar chain is free from the polymer particles 20.

The aromatic amine (Compound A) is not particularly limited, and examples thereof include 2-aminobenzamide, 2-aminobenzoic acid, and 8-aminoindole-1,3,6-trisulphonate. , 8-aminonaphthalene-1,3,6-trisulphonate, 2-amino-9(10H)-acridone, 5-amino fluorescein, dansyl ethylenediamine, 7-amine Base-4-methylcoumarin, 3-aminobenzoic acid, 7-amino-1-naphthol and 3-(ethylamido)-6-amino acridine. Among them, 2-aminobenzamide or 2-aminobenzoic acid is preferred. These compounds are suitably used in terms of the availability of the reagent and the ease of the reaction.

Further, when 2-aminobenzamide or 2-aminobenzoic acid is used as the aromatic amine, 0.35 mol/L is used under normal conditions. However, in this step, the concentration of the aromatic amine in the solution in the well 101 containing the compound containing liquid 25, that is, the solution containing the re-free sugar chain is preferably set to 0.5 mol/L or more, more preferably Set to 1.4 mol/L or more. Thereby, the efficiency of labeling the sugar chain by the compound A can be improved. However, if the concentration of the aromatic amine becomes 3 mol/L or more, the subsequent sugar chain purification In the step, it is difficult to remove the aromatic amine (compound A) which is not used for the labeling reaction of the sugar chain. Therefore, the concentration of the aromatic amine is preferably set to be 1.4 mol/L or more and 3 mol/L or less.

In addition, when the liquid amount of the compound-containing liquid 25 is defined as the liquid amount of the compound-containing liquid 25 based on the polymer particles 20 contained in the pores 101, the liquid amount is usually such that the polymer particles 20 are immersed. The amount of the liquid is, for example, about 50 μL with respect to 5 mg of the polymer particles 20. However, in the present embodiment, it is preferable to set the liquid amount (volume) of the compound-containing liquid 25 to about 100 μL which is twice the amount of the usual liquid amount. Thereby, the efficiency of labeling the sugar chain by the compound A can be improved. However, although the amount of liquid may exceed 100 μL, if it exceeds a certain amount, in the subsequent sugar chain purification step, there is an aromatic amine (compound A) which is difficult to remove the labeling reaction which is not used for the sugar chain. Hey. Therefore, the optimum amount of liquid is set between 100 μL and 200 μL.

More specifically, in the case where 2-aminobenzamide is used as the aromatic amine, it is added to the pores 101 to become 1.4 M 2-aminobenzamide, 1 M sodium cyanoborohydride. 100 μL of a solution obtained by dissolving in a concentration of 30% acetic acid/dimethyl sulfonium (DMSO, Dimethyl Sulfoxide) as a compound-containing liquid 25 was used.

[9-4'] Then, the heater 61 is operated to heat the compound-containing liquid 25, whereby the compound-containing liquid 25 is maintained at a fixed temperature range. Thereby, the sugar chain re-freed from the polymer particles 20 reacts with the compound A, and as a result, the sugar chain is labeled with the compound A.

The pH of the reaction solution (compound containing liquid 25) at this time is preferably 2 to 9, more Preferably, it is 2 to 7, and more preferably 2 to 6.

The temperature of the compound-containing liquid 25 at the time of labeling is preferably maintained at about 0 to 100 ° C, more preferably at about 4 to 95 ° C, and still preferably at a temperature of about 30 to 90 ° C. set up.

Further, when the reaction time is set to the above temperature range, it is usually set to about 0.1 to 20 hours, preferably about 0.6 to 12 hours.

More specifically, when 2-aminobenzamide is used as the aromatic amine, the compound-containing liquid 25 is heated in a temperature range of 30 to 70 ° C for about 1 to 10 hours to carry out a reaction.

The sugar chain was reliably labeled with Compound A by labeling under the above conditions.

The sugar chain is also labeled with Compound A by the steps [9-1'] to [9-4'] as described above.

In the above, the sugar chain purification device and the sugar chain purification method of the present invention have been described with reference to the embodiments of the present invention. However, the present invention is not limited thereto, and each of the components constituting the sugar chain purification device can be configured to have the same function. Replacement is performed. Further, any constituent may be added.

Further, the nozzle of the dispensing mechanism is fixed to the nozzle head in each of the above embodiments, but the invention is not limited thereto, and the nozzle can be detachably attached to the nozzle head.

Further, in the above embodiments, the moving mechanism is configured to move the nozzle of the dispensing mechanism with respect to the plate assembly on the stage. However, the moving mechanism is not limited thereto, and the moving table may be moved with respect to the nozzle. The structure of the board assembly.

Further, the sugar chain purification device may omit the heating mechanism.

Further, in the above-described embodiments, the sugar chain purification apparatus is such that the lower side of the filter which is provided in each of the holes of the upper side plate (the first upper side plate and the second upper side plate) is higher than the upper side in the plate assembly. The negative pressure is such that the liquid passes through the filter. However, the liquid is not limited thereto, and the upper side of the filter may be brought into a positive pressure to pass the liquid through the filter.

Further, the exchange of the first upper side plate and the second upper side plate of the plate assembly and the exchange of the first lower side plate and the second lower side plate may not be performed on the stage.

Further, in each of the above embodiments, the movement of the plate, the disassembly of the plate assembly, and the attachment and detachment of the member are performed by a manual operation of the operator (operator) of the purification device, but the present invention is not limited thereto, and for example, Automate operations using mechanisms such as robotic arms.

Further, the sugar chain purification device may be entirely housed in a chamber that can be internally heated.

[Industrial availability]

The invention provides a sugar chain purification device. This sugar chain purification device includes a plate assembly used for purifying the sugar chain from a solution containing a sugar chain, a stage on which the plate assembly is placed, a suction liquid, and assembly of the plate on the stage. a dispensing mechanism of the nozzle for discharging the body, and a moving mechanism for moving the plate assembly on the stage relative to the nozzle. The plate assembly includes: a first upper side plate, a second upper side plate, a first lower side plate, a second lower side plate, and a support body, wherein the plate assembly system is formed on the stage with respect to the one support body The first upper side plate is exchanged with the second upper side plate, and the first lower side plate is exchanged with the second lower side plate; The first upper side plate has a plate shape, and has a plurality of first holes formed of a first concave portion having an open upper surface and a first through hole penetrating the bottom portion of the first concave portion, and each of the first holes is covered The first through hole is provided in the first filter of the bottom portion of the first recess; the second upper side plate has a plate shape, and has a second recess formed by the upper surface and is formed to penetrate the second through hole. a plurality of second holes of the second through hole at the bottom of the recess and a second hole that covers the bottom of each of the second holes, and is different from the first filter a filter, wherein the first lower side plate has a plate shape and has one first storage portion formed by a third recess having an open upper surface; the second lower side plate has a plate shape and has a fourth opening from the upper surface. a plurality of second storage portions formed by the recesses; wherein the one support system is one of the first upper side plate and the second upper side plate, and one of the first lower side plate and the second lower side plate The person is placed in a state of being assembled up and down and supported. Thereby, the sugar chain can be separated and purified easily and with high precision, and the sugar chain can be fluorescently labeled with good yield. Therefore, the present invention has industrial applicability.

1‧‧‧Sugar chain purification unit (purification unit)

3‧‧‧ stage

4‧‧‧Dispensing agency

5‧‧‧Mobile agencies

6, 6A, 6B‧‧‧ heating mechanism

7‧‧‧cleaning tank

9a, 9b‧‧‧ internal space

10‧‧‧ board assembly

11‧‧‧Control panel

12‧‧‧ wall

13‧‧‧ suction pump

14‧‧‧ tube

20‧‧‧ polymer particles

21‧‧‧ pure water

22‧‧‧Particle dispersion

23‧‧‧Sugar chain containing liquid (solution)

24‧‧‧ cleaning solution

25‧‧‧Compound containing liquid

26‧‧‧Solution solution

27‧‧‧ Non-aqueous solvent

28‧‧‧ water

31‧‧‧ Board assembly placement

32‧‧‧1st upper side panel loading section

33‧‧‧2nd upper side panel loading section

34‧‧‧1st lower side panel mounting section

35‧‧‧2nd lower side panel mounting section

36‧‧‧ Cleaning tank placement

37‧‧‧Processing Liquid Loading Department

41‧‧‧Nozzles (pipettes)

42‧‧‧ pump

43‧‧‧ tube

44‧‧‧Nozzle head

51‧‧‧x-axis moving mechanism (horizontal moving mechanism)

52‧‧‧Y-axis direction moving mechanism (horizontal moving mechanism)

53‧‧‧z axis direction moving mechanism (vertical direction moving mechanism)

61‧‧‧heater

62‧‧‧ chamber

63‧‧‧heater

64‧‧‧ chamber body

65‧‧‧

66‧‧‧Heating device body

67‧‧‧heater

71‧‧‧cleaning fluid

81‧‧‧Multiwell plate

82, 83, 84, 85, 86, 87 ‧ ‧ storage tanks

100‧‧‧1st upper side plate (porous filter plate)

100'‧‧‧2nd upper side panel (cleaning board)

101‧‧‧ holes (1st hole)

101'‧‧‧ hole (2nd hole)

103, 103'‧‧‧ bottom

104, 104'‧‧‧through holes

105‧‧‧Filter (1st filter)

106‧‧‧Filter (2nd filter)

107‧‧‧Protruding

108, 108'‧‧‧ upper surface

109, 109'‧‧‧ lower surface

111‧‧‧LCD screen

200‧‧‧2nd lower side plate (multiwell plate)

201‧‧‧ hole (2nd Reserving Department)

202‧‧‧ Hole Formation Department

203‧‧‧ bottom

204‧‧‧Upper surface

205‧‧‧ Frame Department

300‧‧‧1st lower side panel (waste tray)

301‧‧‧ recessed (first storage department)

302‧‧‧Upper surface

303‧‧‧ recessed part

304‧‧‧ Frame Department

400‧‧‧1st support table (upper support member)

401‧‧‧ bottom

402‧‧‧ openings

403‧‧‧Upper side wall

404‧‧‧Bottom outer wall

405, 406‧‧ ‧ recess

407‧‧‧Washers (sealing members)

408‧‧‧ openings

411‧‧‧ front opening

500‧‧‧2nd support table (lower support member)

501‧‧‧ bottom

502‧‧‧Protruding

503‧‧‧ outer wall

505‧‧‧ recess

506‧‧‧through holes

507‧‧‧Washers (sealing members)

600‧‧‧Adjustment plate (spacer plate)

641‧‧‧ mouth

661‧‧‧ recess

700‧‧‧Support

811‧‧‧ hole

d ₁ , d ₂ , d ₃ , d ₄ ‧‧‧ spacing

Fig. 1 is a perspective view showing a first embodiment of the sugar chain purification device of the present invention.

Fig. 2 is a perspective view showing a plate assembly provided in the sugar chain purification device shown in Fig. 1;

Fig. 3 is an exploded perspective view of the board assembly shown in Fig. 2.

Fig. 4 is a cross-sectional view taken along line B-B of Fig. 2 (a cross-sectional view showing a first state of the plate assembly).

Fig. 5 is a cross-sectional view taken along line B-B of Fig. 2 (a cross-sectional view showing a second state of the plate assembly).

Fig. 6 is a cross-sectional view taken along line B-B of Fig. 2 (a cross-sectional view showing a third state of the plate assembly).

Fig. 7 is a cross-sectional view taken along line B-B of Fig. 2 (a cross-sectional view showing a fourth state of the plate assembly).

Fig. 8 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 9 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 10 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 11 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Fig. 12 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Fig. 13 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Figure 14 is a cross-sectional view showing the steps of purifying the sugar chain by the sugar chain purification apparatus shown in Figure 1 (the cross section observed from the direction of the arrow A in Figure 1) Figure).

Fig. 15 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Fig. 16 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 17 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 18 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Fig. 19 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 20 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view seen from the direction of arrow A in Fig. 1).

Fig. 21 is a cross-sectional view showing a step of purifying a sugar chain by the sugar chain purification apparatus shown in Fig. 1 (a cross-sectional view as seen from the direction of arrow A in Fig. 1).

Fig. 22 is a perspective view showing a second embodiment of the sugar chain purification device of the present invention.

Fig. 23 is a perspective view showing a third embodiment of the sugar chain purification device of the present invention.

1‧‧‧Sugar chain purification unit (purification unit)

3‧‧‧ stage

4‧‧‧Dispensing agency

5‧‧‧Mobile agencies

6‧‧‧heating mechanism

7‧‧‧cleaning tank

10‧‧‧ board assembly

11‧‧‧Control panel

12‧‧‧ wall

13‧‧‧ suction pump

14‧‧‧ tube

22‧‧‧Particle dispersion

23‧‧‧Sugar chain containing liquid (solution)

24‧‧‧ cleaning solution

25‧‧‧Compound containing liquid

26‧‧‧Solution solution

27‧‧‧ Non-aqueous solvent

28‧‧‧ water

31‧‧‧ Board assembly placement

32‧‧‧1st upper side panel loading section

33‧‧‧2nd upper side panel loading section

34‧‧‧1st lower side panel mounting section

35‧‧‧2nd lower side panel mounting section

36‧‧‧ Cleaning tank placement

37‧‧‧Processing Liquid Loading Department

41‧‧‧Nozzles (pipettes)

42‧‧‧ pump

43‧‧‧ tube

44‧‧‧Nozzle head

51‧‧‧x-axis moving mechanism (horizontal moving mechanism)

52‧‧‧Y-axis direction moving mechanism (horizontal moving mechanism)

53‧‧‧z axis direction moving mechanism (vertical direction moving mechanism)

61‧‧‧heater

71‧‧‧cleaning fluid

81‧‧‧Multiwell plate

82, 83, 84, 85, 86, 87 ‧ ‧ storage tanks

100‧‧‧1st upper side plate (porous filter plate)

100'‧‧‧2nd upper side panel (cleaning board)

101‧‧‧ holes (1st hole)

101'‧‧‧ hole (2nd hole)

111‧‧‧LCD screen

200‧‧‧2nd lower side plate (multiwell plate)

201‧‧‧ hole (2nd Reserving Department)

202‧‧‧ Hole Formation Department

203‧‧‧ bottom

300‧‧‧1st lower side panel (waste tray)

301‧‧‧ recessed (first storage department)

400‧‧‧1st support table (upper support member)

411‧‧‧ front opening

500‧‧‧2nd support table (lower support member)

506‧‧‧through holes

700‧‧‧Support

811‧‧‧ hole

Claims (25)

A sugar chain purification device comprising a plate assembly used for purifying the sugar chain from a solution containing a sugar chain, a stage on which the plate assembly is placed, a liquid to be pumped, and the carrier a dispensing mechanism of the nozzle for ejecting the plate assembly on the stage, and a moving mechanism for moving the plate assembly on the stage relative to the nozzle, and the plate assembly includes a first upper side plate a plate shape having a plurality of first holes formed by a first recess opened on the upper surface and having a first through hole penetrating the bottom of the first recess, and the first through hole covering each of the first holes The first filter is provided in the bottom portion of the first recess, and the second upper plate has a plate shape and has a second recess formed by the upper surface and is formed with a second through the bottom of the second recess. a plurality of second holes of the through hole and a second filter that is provided on the bottom of the second recess and that is different from the first filter so as to cover the second through holes of the second holes; 1 lower side plate, which is plate-shaped and has a top a first storage portion formed by a third recessed portion having a surface opening; and a second lower side plate having a plate shape and having a plurality of second storage portions including a fourth concave portion having an open upper surface; and one a support body in which one of the first upper side plate and the second upper side plate and one of the first lower side plate and the second lower side plate are placed up and down and assembled; And the above-mentioned board assembly system is on the above-mentioned stage with respect to the above one support The first upper side plate is exchanged with the second upper side plate, and the first lower side plate and the second lower side plate are exchanged. A sugar chain purification device according to claim 1, further comprising a heating means for heating the plate assembly. The sugar chain purification device according to claim 2, wherein the operation of the heating means is stopped when the first lower side plate and the second lower side plate are exchanged. The sugar chain purification device according to claim 2, wherein the plate assembly is in a first state in which the first upper side plate is supported by the support body and supports the first lower side plate, and the second state is The first lower side plate in the first state is exchanged with the second lower side plate, and the second lower side plate is supported by the support body; and the third state is the first one in the second state. The upper side plate is exchanged with the second upper side plate, and the second upper side plate is supported by the support body, and the second lower side plate is exchanged with the first lower side plate, and the first lower side plate is supported by the support body; In the fourth state, the first lower side plate and the second lower side plate are exchanged in the third state, and the second lower side plate is supported by the support. The sugar chain purification device according to claim 4, wherein in the first state, each of the first holes is in communication with the first storage portion via the first through hole; In the second state, each of the first holes communicates with each of the second storage portions via the first through holes; and in the third state, the second holes pass through the second through holes Further, in the fourth state, each of the second holes communicates with each of the second storage portions via the second through holes. The sugar chain purification apparatus according to claim 5, wherein the sugar chain purification apparatus comprises the following steps: the first step of supplying the sugar chain specific to each of the first pores a second capture step of dispensing the solution from the nozzle into each of the first holes, contacting the solution with the capture carrier, and capturing the sugar chain onto the capture carrier; a third step of removing the substance other than the sugar chain bound to the capture carrier from the pore; the fourth step of re-freeing the sugar chain bound to the capture carrier; and the fifth step Purifying the above-mentioned sugar chain from the above-described capture carrier and purifying it; and in the first step, the second step, the third step, and the fourth step, the plate assembly is in the first state, In the fifth step, the plate assembly is sequentially set to the second state, the third state, and the fourth state. The sugar chain purification device of claim 6, wherein the heating mechanism is In the first step, the second step, the third step, the fourth step, and the fifth step, the heating temperature of the panel assembly or the first upper plate may be changed. The sugar chain purification device according to claim 2, wherein the heating means has a heater embedded in the stage. The sugar chain purification device of claim 2, wherein the heating mechanism is a thermostat having a heater. The sugar chain purification device according to claim 1, wherein the support body has an upper support member that supports the one of the first upper side plate and the second upper side plate, and is configured separately from the upper support member to support the first a lower support member of the one of the lower side plate and the second lower side plate, and the upper support member and the lower support member support the first upper side plate and the second upper side plate by the upper side support member One of the first lower side plates and the second lower side plate is detachably/assembled in a state in which the lower support member supports the one of the first lower side plate and the second lower side plate. The sugar chain purification device according to claim 10, wherein the exchange of the first upper side plate and the second upper side plate and the exchange of the first lower side plate and the second lower side plate respectively form the upper support member and the lower side The side support member is dismantled. The sugar chain purification device according to claim 1, wherein in the plate assembly, the lower surface of the one of the first upper side plate and the second upper side plate and the first lower side plate and the second lower portion are One of the above side panels The upper surfaces are spaced apart and have a pitch of 0.5 mm or more and 10 mm or less. The sugar chain purification device according to claim 1, wherein the first filter is composed of a porous film or a nonwoven fabric. The sugar chain purification device according to claim 13, wherein the mesh of the first filter is 0.1 to 50 μm. The sugar chain purification device according to claim 1, wherein the second filter is composed of silicone. The sugar chain purification device according to claim 1, wherein the dispensing mechanism has a pump and a tube connecting the pump to the nozzle. The sugar chain purification device according to claim 1, wherein the moving mechanism has a horizontal direction moving mechanism that moves the nozzle in a horizontal direction and a vertical direction moving mechanism that moves the nozzle in a vertical direction. The sugar chain purification device according to claim 1, wherein the first upper side plate mounting portion on which the plurality of the first upper side plates are stacked and placed on the stage is provided, and the plurality of the second upper side plates are overlapped and carried The second upper side plate mounting portion, the first lower side plate mounting portion on which the plurality of first lower side plates are stacked, and the second lower side plate on which the plurality of second lower side plates are stacked and placed Set up. The sugar chain purification device according to claim 1, further comprising a cleaning tank for cleaning the nozzle. A method for purifying a sugar chain, which comprises the step of purifying the sugar chain using the sugar chain purification device of claim 6, and in the fourth step, the sugar chain bound to the capture carrier Freed again and the sugar chain was labeled with a labeling reagent. The method for purifying a sugar chain according to claim 20, wherein in the fourth step, after the sugar chain is re-free from the capture carrier, the sugar chain is labeled with the labeling reagent. The method for purifying a sugar chain according to claim 21, wherein the labeling reagent is a fluorescent substance composed of an aromatic amine. The method for purifying a sugar chain according to claim 22, wherein the concentration of the fluorescent substance in the solution containing the sugar chain re-freed is 0.5 mol/L or more. The method of purifying a sugar chain according to claim 22, wherein the fluorescent substance is selected from the group consisting of 2-aminobenzamide, 2-aminobenzoic acid, 8-aminoindole-1,3,6-trisulphonate , 8-aminonaphthalene-1,3,6-trisulphonate, 2-amino-9(10H)-acridone, 5-amino fluorescein, dansyl ethylenediamine, 7-amine At least one of ketomethyl coumarin, 3-aminobenzoic acid, 7-amino-1-naphthol, and 3-(acetamido)-6-aminopyridinium. The method of purifying a sugar chain according to claim 20, wherein the capture carrier is a polymer particle having a mercapto group or an oxyamino group.