JP2006247535A - Method for promoting reaction by using rotating magnetic field - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for promoting a reaction by using a new rotating magnetic field, by which a specific component can be detected with high sensitivity, the concentration of the specific component can be measured, the reaction time can be shortened remarkably and the amounts of reagents to be used can be saved. <P>SOLUTION: The method for promoting the reaction by using the rotating magnetic field comprises the steps of: setting the ratio of the horizontal component of a magnetic field to the vertical component; arranging at least one or more magnets on the upper or lower outside of a reactor; rotating the arranged magnets to generate the rotating magnetic field, to rotate magnetic beads packed in the reactor and to form a bar-shaped magnetic bead cluster aligned in the direction of the magnetic field; reacting the specific component in a sample solution introduced into the reactor with an active component of magnetic beads; and separating a reaction product from the magnetic bead cluster by the flow of the sample solution to be generated by the rotating magnetic field. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a reaction promoting method using a rotating magnetic field.

  In recent years, microanalysis such as biotechnology and environmental analysis has been important and required to be performed quickly, and as a result, research and development of microanalysis chips has become a central part of portable analytical instruments. It is being advanced. In addition, research on synthesis reactions using microreactors produced by microfabrication technology is also underway.

  Conventionally, when analyzing trace components in liquids, the reaction of elements that detect light and electrical signals is expected to be fast and highly practical, but the components to be analyzed diffuse in the dispersion medium. In addition, since the progress of the process of collision and reaction with the catalyst, antibody, enzyme, etc. at the molecular level is slow, it has been the cause of the long analysis time. For example, ELISA (enzyme-labeled immunization) is used to test mad cow disease, but this method requires several hours or more for the reaction time of the antibody-antigen reaction, so it takes a long time to understand the results. It was a big problem.

  In addition, in the conventional biochemical analysis method, a microwell plate or the like having a plurality of wells having a radius and depth in millimeter units called microwells is used, and a liquid sample of the order of 100 μl is used in this well. And is allowed to react with the antibody or the like immobilized on the wall of the well in advance. However, there is a problem that it takes a long time of around several hours for the binding molecules in the sample solution to almost complete the binding reaction with the antibody immobilized on the well wall.

Therefore, a method for shortening the reaction time has been proposed (Patent Document 1) in which an antibody or the like is immobilized on the surface of the magnetic beads and the antibody-immobilized magnetic beads are reacted with a sample containing a reaction product. In this method, a substance to be measured is immobilized in a predetermined measurement container in advance, and an antibody-immobilized marker magnetic bead in which an antibody that binds to the substance to be measured is immobilized on the magnetic bead is reacted. The reaction is completed in a short time by applying a magnet to increase the sedimentation rate of the antibody-immobilized marker magnetic beads. When the substance to be measured in the sample reacts with the antibody, the antibody-immobilized marker magnetic beads and the substance to be measured are bound by the antibody-antigen reaction, and the substance to be measured is immobilized throughout the predetermined container. For this reason, a sedimentation pattern that spreads uniformly is formed, and the substance to be measured present in the sample can be determined with high sensitivity.
Patent No. 2614997

  However, the method described in Patent Document 1 is mainly intended to determine the presence or absence of the target substance to be measured in the sample, and does not consider shortening of the reaction time. In other words, stirring and reaction are stopped each time, and the magnetic beads are attracted to the wall of the test tube (reactor) with a magnet to separate the product in the supernatant solution (that is, the substance to be measured). There was still a problem that the reaction (analysis) time was long. Further, in this case, there are problems in the sensitivity of detection and measurement, and in saving the amount of reagent used.

  Therefore, the present invention was made in view of the circumstances as described above, solves the conventional problems, can significantly reduce the reaction time, and can also save the amount of reagents used, It is an object to provide a reaction promotion method using a new rotating magnetic field.

According to the present invention, as a means for solving the above problems, first, a rotating magnetic field is applied to a magnetic bead on which an active component that reacts with a specific component in a sample solution is fixed. A reaction promotion method using a rotating magnetic field that promotes a reaction, which comprises the following steps:
<1> At least one or more above or below the reactor, which is a reaction field between a specific component and an active component constructed on a planar substrate, by setting a ratio between a horizontal component and a vertical component of the magnetic field Arranged magnets;
<2> A rotating magnetic field is generated by rotating a magnet, whereby the magnetic beads filled in the reactor are rotated and formed as rod-shaped magnetic bead clusters aligned in the magnetic field direction. Converge to the neighborhood;
<3> reacting a specific component in the sample solution introduced into the reactor with the active component of the magnetic beads;
<4> The product after the reaction is separated from the magnetic bead cluster by the flow of the sample solution by the rotating magnetic field;
It is characterized by including.

  According to the present invention, secondly, the magnet is arranged using four magnets, each being symmetrically arranged around the central axis of the rotating magnetic field, and thirdly, The horizontal / vertical component ratio of the magnetic field is 1/3 or less, and fourth, the magnet is a permanent magnet.

  And fifth, the present invention is characterized in that the surface of the magnetic beads is coated with a polymer component having affinity for biomolecules.

  According to the present invention as described above, a specific component can be detected with high sensitivity and the concentration thereof can be measured, the reaction time can be greatly shortened, and the amount of reagents used can be saved.

  The present invention has the features as described above, and the embodiments thereof will be described in detail below.

The present invention is a reaction promotion method using a rotating magnetic field in which a rotating magnetic field is applied to a magnetic bead on which an active component that reacts with a specific component in a sample solution is fixed to accelerate the reaction between the specific component and the active component. And at least the following steps. Ie:
<Step 1> The ratio between the horizontal component and the vertical component of the magnetic field is set, and above or below the reactor that is the reaction field between the specific component and the active component constructed on the planar substrate (that is, the reactor At least one or more magnets are arranged above or below when a sample solution containing magnetic beads is placed horizontally. Usually, the magnet is hold | maintained at the magnet holder installed in the fixed distance d rather than the reactor, as illustrated in FIG. This distance d can be set to 1 to 10 mm, for example, whereby an efficient reaction promoting effect can be obtained, but is particularly preferably set to 1 to 5 mm. In the case of using one magnet, in order to sufficiently obtain the effect of the present invention, it is preferable to dispose the magnet away from the central axis (rotating axis) of the rotating magnetic field in Step 2. Furthermore, this reactor is characterized by having a high reagent-saving effect that can be carried out even in a reaction of μl order by using a micro reactor.
<Step 2> Next, a magnetic field is generated by rotating the magnet holder (that is, the magnet), thereby rotating the magnetic beads filled in the reactor, and as illustrated in FIG. It is formed as a rod-shaped magnetic bead cluster aligned in the magnetic field direction, and converges near the central axis of the rotating magnetic field. In order to sufficiently obtain the effects of the present invention, the central axis of the rotating magnetic field is preferably matched with the central axis of the reactor.
<Step 3> The specific component in the sample solution introduced into the reactor is reacted with the active component of the magnetic beads.
<Step 4> The product after the reaction between the specific component and the active component is separated from the magnetic bead cluster by the flow of the sample solution by the rotating magnetic field. At this time, the magnetic bead cluster is maintained in the state of being held on the central axis of the rotating magnetic field.

  Furthermore, the arrangement of the magnets in step 1 will be described. As shown in the example of FIG. 1, four magnets are used, and each of the four magnets is arranged symmetrically around the central axis of the rotating magnetic field. Is preferred. That is, they are arranged at an equal distance of 90 ° from the central axis of the rotating magnetic field. And in order to exhibit the effect of this invention more efficiently, it is more preferable to arrange | position a magnet so that a non-homogeneous magnetic field may be generated in consideration of the polarity (N pole, S pole) which a magnet has. Specifically, for example, the four magnet arrangements may be arranged asymmetrically in the polarity of the magnets, that is, “NNNS arrangement” or “SSSN arrangement”, where N and S are closer to the reactor. The magnet polarity (N pole or S pole) is shown. In other words, in the “NNNS arrangement”, the magnets are arranged so that the N pole is on the upper side, the N pole is on the left side, the N pole is on the lower side, and the S pole is on the right side when viewed from the center of the reactor. In the case of “SSSN arrangement”, the magnet is arranged so that the S pole is on the upper side, the S pole is on the left side, the S pole is also on the lower side, and the N pole is on the right side. By arranging the magnets in this way, the magnetic beads are aggregated in a disk shape, and the magnetic beads are maintained at the center of the rotating magnetic field (that is, the center of the reactor) even when the sample solution is flowed. The product after the reaction is separated from the magnetic bead cluster by the flow of the sample solution by the rotating magnetic field. As a result, the specific component can be detected with high sensitivity and the concentration thereof can be measured, the reaction time can be greatly shortened, and the amount of reagents used can be saved.

  The shape of the magnet is not particularly limited in order to arrange the polarities as described above asymmetrically, but it is convenient to have a rod shape (for example, a rod magnet having a diameter of 1 to 2 mm and a length of 10 mm). Is preferable.

  In the present invention, the ratio of the horizontal component / vertical component of the magnetic field is preferably 1/3 or less (horizontal component / vertical component of magnetic field ≧ 1/3). At this time, the arrangement in which the horizontal component is stronger than the vertical component is more effective in terms of promoting the reaction. In the present invention, the horizontal component is the horizontal component of the magnetic field with respect to the substrate, and the vertical component is also the vertical component of the magnetic field with respect to the substrate. In the present invention, the horizontal and vertical components, and the strength of the magnetic field vary depending on the installation location of the reactor with respect to the magnetic field.

  The magnet used in the present invention is preferably a permanent magnet. The “permanent magnet” is a ferromagnetic material having a coercive force having a residual magnetic flux in which about 50 to 80% of magnetization remains even when the magnetic field is zero. The density of the “residual magnetic flux” is not particularly limited. For example, the density of the magnetic flux density is 0.01 to 0.3 Tesla.

  In the present invention, the reaction can be promoted by generating a homogeneous magnetic field using an electromagnet as the magnet. An "electromagnet" is a coil around a soft iron core, and a magnetic pole of an appropriate shape is installed at the tip of the iron core. The magnet is magnetized when the current is passed, and the magnetization disappears when the current is turned off. It is a magnet and there is a residual magnetic flux. The intensity of the generated magnetic field in this electromagnet is proportional to the product of the number of turns of the coil and the current, and is also affected by the shape and size of the iron core and magnetic pole.

  The magnetic bead cluster in the case of using this electromagnet will be described. The relationship between the magnetic field strength and the magnetic field rotation frequency is arranged by a dimensionless number Ma called Mason number (the following equation (I)).

(Where μ ₀ is the permeability of vacuum, η is the viscosity of the dispersion medium (sample solution), ω is the rotational speed of the magnetic field (radian / s), and M is the magnetization of the magnetic particles)
Considering the magnetic bead cluster based on this formula, when the magnetic bead volume fraction is low (10 ^{−4 to} 0.02), Ma1 becomes a branch point regardless of the fraction. When Ma ≧ 1, the shape of the rod-shaped magnetic bead cluster (chain cluster) is maintained, but when Ma ≦ 1, the rod-shaped magnetic bead cluster (chain cluster) is decomposed into particles and isotropic clusters (spherical) Reorganized to a close shape. Furthermore, at high volume fractions, the rod-like magnetic bead clusters aggregate and become thick, and the cluster structure and dynamic behavior change. When the frequency of the external field increases, the polarization is delayed and the cluster is formed in a disk shape.

Here, a magnetic bead cluster formed by a direct current magnetic field is illustrated in FIG. As shown in FIG. 3, in the DC magnetic field, rod-like magnetic bead clusters (chain clusters) aligned in the magnetic field direction are formed (FIG. 3a). When the rotation frequency is as low as 0.1 Hz (Ma = 1.3 × 10 ^-3 ), the rod-shaped magnetic bead cluster (chain cluster) rotates following the rotating magnetic field, and the volume fraction of magnetic beads is high. Some rotating clusters collide with each other to form thick clusters (FIG. 3b). On the other hand, when the rotating magnetic field frequency reaches 2 Hz (Ma = 2.5 × 10 ⁻² ), the rod-like magnetic bead clusters are decomposed and shortened, and finally aggregate to form a disk-like cluster (FIG. 3c). Further, at 10 Hz (Ma = 0.13), it was further aggregated (FIG. 3d). On the surface of the disk-like cluster, the magnetic beads form a short rod-like magnetic bead cluster, which moves while rotating on the surface by an external magnetic field.

  Thus, when a high-density magnetic cluster (magnetic colloid) is placed in a homogeneous magnetic field and the magnetic field is rotated to generate a rotating magnetic field, first, rod-shaped magnetic bead clusters are formed and rotated. This can increase the collision frequency between the active component immobilized on the magnetic bead cluster and the specific component in the sample solution, thereby promoting the reaction. The rod-like magnetic bead clusters exert an attractive force to each other, and are formed into disk-like clusters as described above. The particles on the surface form short rod-like magnetic bead clusters and circulate while rotating on the surface of the disk-like clusters. As a result of the rotation of the rod-like magnetic bead cluster, the disk-like cluster also rotates slowly. In this case, the reaction is promoted at the surface portion of the disk-like cluster. This Mason number can also be applied to permanent magnets.

  The “specific component” in the present invention is a substance that is a target of various reactions in a sample solution. For example, albumin, prostaglandins, steroid hormones and various immune antibodies (IgG, IgA, IgM , IgE, etc.), various amino acids (glutamic acid, leucine, phenylalanine, valine, alanine, γ-aminobutyric acid, etc.) and various chemical substances. In addition, in the method of this application, “reaction in a specific component” for which an accelerating effect is obtained means a concentration measurement reaction, a substance detection reaction, a catalyst, etc. using an antibody-antigen reaction of a target substance in a sample solution. It means various reactions such as a chemical synthesis reaction utilizing a reaction and a separation reaction of a specific component utilizing the magnetism of magnetic beads. An “active ingredient” is a substance that reacts with the specific ingredient. For example, catalytic substances (for example, various enzymes such as protein kinases, tyrosine kinases, and MAP kinases) having an effect of promoting biological reactions and chemical reactions, various antibodies (anti-albumin antibodies, anti-prostaglandin antibodies, anti-steroid hormone antibodies, And biological substances such as anti-IgG antibody, anti-IgA antibody, anti-IgM antibody, anti-IgE antibody) and various chemical substances. The magnetic beads on which the active ingredient is immobilized are preferable because the reaction can be accelerated more greatly by being added in excess of the amount of the specific ingredient.

  The “magnetic beads” are not particularly limited as long as they are insoluble in an aqueous solution and exhibit magnetism. Conventionally used ferromagnetic beads may be used, but superparamagnetic fine particles are more preferable because the magnetic dipole becomes zero when the external magnetic field becomes zero, and magnetic aggregation due to magnetism does not occur. Then, by applying a magnetic field to this superparamagnetic bead, a rod-shaped magnetic bead cluster is formed. By applying a magnetic field to this rod-shaped magnetic bead cluster, the magnetic bead cluster is rotated and fixed to the above-mentioned specific component and magnetic bead. The collision frequency with the activated active ingredient is increased, and as a result, various reactions are promoted. The particle size of the magnetic beads is not particularly limited. For example, the particle size is preferably 10 μm to 1 μm, and the particle size of the magnetic beads may be on the order of nm.

  In addition, when the specific component is a biomolecule, the surface of the magnetic beads may be coated with a polymer component having an affinity for the biomolecule in order to further promote the binding with the specific component. It can be appropriately selected according to the purpose, conditions and the like. Examples of the “polymer component” include various proteins such as avidin that specifically bind to biotin, nucleic acids such as DNA and RNA, and epoxy groups, tosyl groups, amino groups, carboxyl groups, and the like that have high binding properties to proteins. Various active groups etc. are mentioned.

  As a method for immobilizing each of the active ingredients on the magnetic beads, any method such as physical adsorption or formation of a chemical covalent bond can be appropriately employed. For active ingredients with high physical adsorption ability such as antibodies and high molecular weight proteins, immobilization by physical adsorption is preferable, and for active ingredients with low physical adsorption ability such as hormones, chemical covalent bonds Immobilization by is preferred. Many methods of immobilization have already been proposed, and an immobilization method may be adopted from known methods according to the type and characteristics of the active ingredient to be immobilized.

  In the present invention, the concentration of a specific component in a sample solution and the presence thereof can be detected by binding a labeling substance directly to the active component or magnetic beads and analyzing the labeling substance. As the “labeling substance”, various fluorescent substances, chemiluminescent substances, enzymes and the like can be used. For example, as fluorescent substances, green fluorescent protein, red fluorescent protein, chlorophyll, reserpine, folic acid, etc., and derivatives thereof can be used. As chemiluminescent substances, luminol, indole, lophine, etc., and as enzymes, β -D galactosidase, alkaline phosphatase, etc. can be used.

  In the reaction promotion method of the present invention, the migration effect of individual magnetic beads or / and magnetic bead clusters may be used based on the effect of the magnetic field gradient. It means the gradient of the strength that the current affects the other magnets and current around it.

  Further, the magnet holder is driven by a motor (driving machine). The “motor” is limited in its size and the like as long as it can generate power using electricity or the like as an energy source and can rotate a member such as a magnet holder that holds the magnet by the power. It is not something. For example, a small motor (manufactured by Mabuchi Corporation: rotational speed 7800 rpm, 130 revolutions / second) can be used. Of course, this motor can have more than one motor to get more power.

  Hereinafter, the reaction promoting method of the present invention will be described in more detail and specifically with reference to examples. Of course, the present invention is not limited to the following examples.

(Example 1) Design of microreactor (reaction chamber) and examination of stirring efficiency of magnetic beads (1) Creation of microreaction chamber
A chromium photomask drawn with an EB apparatus was developed into a positive type and irradiated with ultraviolet light from above the SU-8, which is a spin-coated ultraviolet photosensitive resin. Thus, a mold (convex mold) in which the reaction chamber was octagonal was created. Next, PDMS (polydimethylsilicone) was poured into the SU-8 mold and allowed to solidify overnight to create a PDMS mold (concave), with a large Aspect ratio (depth / diameter) and circular cross section The reaction promoting effect of the present invention was examined in a reaction chamber close to.

  Next, as illustrated in FIG. 4a, a type A micro reaction chamber was prepared by pasting PDMS solidified on a preparation. The Aspect ratio of this type A micro reaction chamber was 0.0021.

  Further, as illustrated in FIG. 4b, a B-type micro reaction chamber was created in which a hole was formed in PDMS with a belt punch and the upper and lower sides were sandwiched between preparations. The PDMS thickness was adjusted so that the reaction chamber volume was the same for both A and B types. For the B type, two types were created: a B-L type with a small Aspect ratio (Aspect ratio = 0.29) and a B-H type with a large Aspect ratio (Aspect ratio = 0.81).

In addition, for each of A type, BL type, and BH type, three prototypes were created and examined.
(2) Measurement of reaction rate 5 μl of 0.5 mg / ml primary antibody (Anti-RAT IgG) as an active ingredient was immobilized on magnetic beads at 4 ° C. overnight. A sample solution containing this magnetic bead and 100 ng / ml antigen (RAT IgG) as a specific component is injected into a micro reaction chamber, stirred and bound for 1 minute, and then 0.6 mg / ml secondary antibody (Anti -RAT IgG) 5 μl was immobilized in the reaction chamber for 1 minute. Finally, the substrate (ABTS) was poured into the reaction chamber and stirred for 1 minute at a temperature of 20 ° C. The absorbance of the reaction solution after stirring was measured to confirm the reaction rate. The antigen / antibody / substrate was injected in an amount of 2 to 3 μl according to the volume of the reactor.
(3) Measurement results
In three types of micro reaction chambers (A type, BL type and BH type), the absorbance was measured three times, and the average was obtained.

  The results are shown in Table 1. From the results shown in Table 1, the results of FIG. 5 could be derived. The numerical value in the parenthesis in FIG. 5 is the Aspect ratio of each reaction chamber. From Table 1 and FIG. 5, it was confirmed that the absorbance value (absorbance coefficient) was larger as the Aspect ratio of the reaction chamber was closer to 1, that is, the reaction rate was faster. In the case of a flat reaction chamber with a small Aspect ratio, the horizontal component of the magnetic field is important.

Although not shown in the figure, when the movement of the magnetic beads in the B-type reaction chamber was observed with a CCD camera, the magnetic beads aggregated in the reaction chamber moved effectively without leaking in the rotating magnetic field. It was also confirmed that.
(Example 2) Examination of stirring effect under magnetic field application conditions (1) Reagent and apparatus Magnetic beads Dynabeads MyOne Streptavidin, primary antibody ANTI-RAT IgG biotin Conjugate, antigen IgG from rat serum, secondary antibody ANTI-RAT IgG- POD Conjugate, Substrate ABTS solution, Spectrophotometer Nano Drop (NanoDrop Technologies), Gauss meter (FWBELL.), Magnet A (Neodymium magnet φ10 × 5 4200 Gauss), Magnet B (Neodymium magnet φ2 × 10 3600 Gauss) used.
(2) Measurement of magnetic field strength and measurement of enzyme reaction (A) Magnetic field strength measurement For magnetic field strength measurement, a slide glass was fixed to an XY stage, a stirrer was fixed to an aluminum base, and the magnetic field strength was measured with a gauss meter.

  The measurement results were as shown in the magnetic field vector diagrams shown in FIGS.

  FIG. 6 a shows the magnetic field by magnet A. The central rectangle indicates the magnet, the dotted lines at both ends indicate the rotation axis of the stirrer, and the line on the rotation axis indicates the position of the reactor. What was measured is shown by a solid line. A dotted arrow indicates a line symmetric with respect to the vertical axis.

FIG. 6b shows a magnetic field (arranged NSSS pole) by four magnets B. The two rectangles in the lower part of the figure show the magnet, the right shows the N pole, and the left shows the S pole. Further, the rotation axis indicates the center vertical line, and the base portion of the arrow indicates the distance of the magnet from the bottom to 1, 1.5, 2, 2.5, 5, and 7.5 mm, respectively.
(B) Enzyme reaction measurement In the enzyme reaction measurement, magnetic beads pre-bound to the secondary antibody are placed in a microreactor, ABTS is flowed at a flow rate of 2 μl / min, and the extinction coefficient of the enzyme reaction product every 2 minutes. Was measured.

  The measurement results were as shown in FIGS. 7a and 7b.

FIG. 7a shows the time course of the extinction coefficient when stirring with the magnet A, and FIG. 7b shows the time course of the extinction coefficient when stirring with the four magnets B (NSSS pole arrangement). It was also confirmed that the reaction promoting effect was different depending on the change in the distance d to the stirrer.
(C) Results In designing the stirrer, by adding a horizontal magnetic field vector to the reactor, the dipole has a large horizontal vector and a relatively high extinction coefficient at 1.5 mm and 2.5 mm. It was confirmed that At the NSSS electrode, it was confirmed that at 0 mm, the magnetic beads that could normally be collected could not be collected and the stirring efficiency was high.
(Example 3) ELISA method biosensor using magnetic bead cluster agitation In the enzyme immunolabeling analysis method (ELISA method), magnetic bead clusters are formed, and the magnetic bead clusters are used for agitation, followed by the flow method (continuous Act). In the present invention, the magnetic stirring system is such that the magnetic beads do not flow out even by the flow method.

  Specifically, PDMS was injected into a mold processed by photolithography to create a microreactor (microreactor). The permanent magnet was rotated with a small motor to generate a rotating magnetic field, and the magnetic beads were agitated to promote the reaction.

  The sample solution (reagent) was introduced into the reactor by flowing the reagent at a flow rate of 5 μL / min using a syringe pump / valve system. The reactor wall was subjected to blocking treatment (MPC polymer), antigen RAT-IgG was used, and magnetic beads conjugated to the primary antibody in advance were used.

  As a result, primary antibody-antigen reaction (1 minute) → wash (2 minutes) → antigen-secondary antibody (1 minute) → wash (2 minutes) → enzyme reaction (4 minutes) Became possible. This is because the substrate is fed by the flow method, the product is carried out, and the reaction is effectively promoted by stirring magnetic beads. As an example of the reaction speed, the time course of the product concentration after the substrate injection is shown in FIGS. Based on these results, we were able to establish the basis for reagent-saving and rapid analysis technology suitable for ELISA.

  Of course, the present invention is not limited to the above examples, and it goes without saying that various aspects are possible in detail.

It is the top view and side view which illustrated typically one Embodiment of the reaction promotion apparatus in this invention. It is the figure which illustrated the shape of the rod-shaped magnetic bead cluster in this invention, a is a theoretical simulation figure, b is the expanded sectional view which illustrated typically the inside of the reactor in FIG. In this invention, it is the figure which illustrated the shape of the magnetic bead cluster at the time of using an electromagnet, a is the shape of the rod-shaped magnetic bead cluster in a direct current magnetic field, b is 0.1 Hz (Ma = 1.3 * 10 < ^-3 >). ), The shape of the rod-shaped magnetic bead cluster in c) is the shape of the rod-shaped magnetic bead cluster (formed in a disk shape) at a rotational frequency of 2 Hz (Ma = 2.5 × 10 ⁻² ), and d is the rotational frequency of 10 Hz (Ma = 0.13). ) Shows the shape of the rod-shaped magnetic bead cluster (formed in a disk shape). It is the schematic diagram which illustrated the shape of the micro reaction chamber in this invention, a shows the A type micro reaction chamber and b shows the B type micro reaction chamber. It is the graph which showed the relationship between the Aspect ratio of a reaction chamber, and the reaction promotion effect (absorbance). It is a magnetic force line vector diagram in each of magnet A and magnet B in Example 2, a shows the case of magnet A, and b shows the case of magnet B (four pieces). It is the figure which showed the time passage of the extinction coefficient in each of the magnet A in the Example 2, and the magnet B, a shows the case of the magnet A and b shows the case of the magnet B (4 pieces). It is the graph which showed time course of the product concentration after substrate injection, a shows the relation between an extinction coefficient and enzyme reaction time, and b shows the relation between an extinction coefficient and antigen concentration.

Claims (5)

A reaction promotion method using a rotating magnetic field that applies a rotating magnetic field to a magnetic bead on which an active component that reacts with a specific component in a sample solution is fixed, and promotes the reaction between the specific component and the active component. Process:
<1> At least one or more above or below the reactor, which is a reaction field between a specific component and an active component constructed on a planar substrate, by setting a ratio between a horizontal component and a vertical component of the magnetic field Arranged magnets;
<2> A rotating magnetic field is generated by rotating a magnet, whereby the magnetic beads filled in the reactor are rotated and formed as rod-shaped magnetic bead clusters aligned in the magnetic field direction. Converge to the neighborhood;
<3> reacting a specific component in the sample solution introduced into the reactor with the active component of the magnetic beads;
<4> The product after the reaction is separated from the magnetic bead cluster by the flow of the sample solution by the rotating magnetic field;
A reaction promoting method using a rotating magnetic field.   2. The reaction promotion method according to claim 1, wherein the magnets are arranged using four magnets, each of which is symmetrically arranged around the central axis of the rotating magnetic field.   The method according to claim 1 or 2, wherein the ratio of the horizontal component / vertical component of the magnetic field is 1/3 or less.   The method according to claim 1, wherein the magnet is a permanent magnet. 5. The reaction promoting method according to claim 1, wherein the surface of the magnetic beads is coated with a polymer component having affinity for biomolecules.