Immunoreaction reagent, method for producing same, and immunoreaction reagent kit

Abstract

The present invention addresses the problem of providing an immunoreaction reagent in a freeze-dried state, the reagent being used in an extracorporeal diagnostic agent or the like and having good measurement reproducibility. The problem is solved by obtaining an immunoreaction reagent by: letting solid-phase antibodies for antigens subject to measurement coexist with a defoaming agent in a solution and freeze-drying the solution; letting solid-phase antigens for antibodies subject to measurement coexist with a defoaming agent in a solution and freeze-drying the solution; or letting solid-phase antigens acting as competitors with antigens subject to measurement in bonding with antibodies for the antigens subject to measurement coexist with a defoaming agent in a solution and freeze-drying the solution.

Description

Immune reaction reagent, its manufacturing method and immune reaction reagent kit

[0001] The present invention relates to a freeze-dried immune reaction reagent for use in in vitro diagnostics, a method for producing the same, and an immune reaction reagent kit.

[0002] Antibodies immobilized on solid-phase carriers such as beads or microparticles (hereinafter referred to as solid-phase antibodies), antigens immobilized on solid-phase carriers (hereinafter referred to as solid-phase antigens), antibodies conjugated with labels such as enzymes (hereinafter referred to as labeled antibodies), and antigens conjugated with labels such as enzymes (hereinafter referred to as labeled antigens) are widely used in fields such as clinical testing. However, the activity of antibodies, antigens, enzymes, etc. is easily affected by temperature, making it difficult to store them in a state that maintains their activity for long periods of time. For this reason, various stabilization methods have been proposed to maintain their activity.

[0003] Patent Document 1 reports that labeled antibodies can be stabilized by adding disaccharide or higher non-reducing sugars and sugar alcohols, etc., and then freeze-drying the resulting antibody. Patent Document 2 reports that human alkaline phosphatase can be stabilized by freeze-drying the antibody in the presence of a sugar selected from the group consisting of galactose, lactose, and fructose, and albumin or dextran. However, when freeze-dried immunoreaction reagents are obtained with the storage stability required in the field of clinical testing by selecting appropriate compounds, sugars, proteins, etc., the reagents often become a hard dried cake, which has poor solubility after reconstitution. As a result, measurement reproducibility is poor, hindering highly accurate measurement of the target component.

[0004] JP-A-60-149972 Patent No. 4169344

[0005] In the field of clinical testing, long-term storage stability of various reagents is required. Reproducibility during measurement is also required. When using a freeze-dried reagent for measurement, it must be dissolved in a dissolving solution or the like. However, during dissolution, air bubbles may be generated due to porosity (voids) resulting from freeze-drying. The present inventors have discovered that there is variability in the presence or absence of air bubbles and their size, and that this variability in the presence or absence of air bubbles and their size leads to poor reproducibility. Therefore, an object of the present invention is to provide a freeze-dried immune reaction reagent with good measurement reproducibility for use in in vitro diagnostics, etc., a method for producing the same, and an immune reaction reagent kit.

[0006] As a result of intensive research to solve the above-mentioned problems, the inventors discovered that adding an antifoaming agent to a freeze-dried immune reaction reagent improves measurement reproducibility, and thus completed the present invention.

[0007] That is, the present invention is as follows: (1) An immune reaction reagent comprising: a freeze-dried reagent containing a solid-phase antibody against an antigen to be measured and an antifoaming agent. (2) An immune reaction reagent comprising: a freeze-dried reagent containing a solid-phase antigen against an antibody to be measured and an antifoaming agent. (3) An immune reaction reagent comprising: a freeze-dried reagent containing a solid-phase antigen that competes with the antigen to be measured for binding to the antibody against the antigen to be measured and an antifoaming agent. (4) The immune reaction reagent according to any one of (1) to (3), wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent. (5) A method for producing an immune reaction reagent, comprising: causing a solid-phase antibody against an antigen to be measured and an antifoaming agent to coexist in a solution, followed by freeze-drying. (6) A method for producing an immune reaction reagent, comprising causing a solid-phase antigen against an antibody to be measured and an antifoaming agent to coexist in a solution, followed by freeze-drying. (7) A method for producing an immune reaction reagent, comprising: causing a solid-phase antigen that competes with the antigen to be measured for binding to an antibody against the antigen to coexist in a solution with an antifoaming agent; and freeze-drying the resulting solution. (8) A method for producing an immune reaction reagent according to any one of (5) to (7), wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent. (9) A method for producing an immune reaction reagent according to any one of (5) to (8), wherein the solid-phase antibody or the solid-phase antigen is dispersed as a suspension, and then immediately frozen and freeze-dried. (10) An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antibody against the antigen to be measured and an antifoaming agent; and a reagent containing a labeled antibody capable of binding to the antigen. (11) An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antibody against the antigen to be measured and an antifoaming agent; and a reagent containing a labeled antigen capable of binding to the solid-phase antibody. (12) An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antigen against an antibody to be measured and an antifoaming agent; and a reagent containing a labeled antibody capable of binding to the antigen.(13) An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antigen that competes with the antigen to be measured for binding to an antibody against the antigen and an antifoaming agent, and a reagent containing the solid-phase antigen and a labeled antibody capable of binding to the antigen to be measured. (14) The immune reaction reagent kit according to any one of (10) to (13), wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent.

[0008] The present invention will be described in detail below. The measurement target in the present invention is an antigen or antibody. Antigens are not particularly limited, but include substances used in clinical tests, such as squamous cell carcinoma-associated antigen (SCC antigen), prostate-specific antigen (PSA), thyroid-stimulating hormone (TSH), thyroxine, triiodothyronine, testosterone, estradiol, progesterone, cortisol, aldosterone, homocysteine, β2-microglobulin (BMG), brain natriuretic peptide (BNP), and C-peptide. Among these, aldosterone and homocysteine ​​are preferred. The reason for this is as follows: Specifically, the inventors discovered a rare phenomenon in which, particularly with this antigen, measurement values ​​fluctuate due to interference from air bubbles within the measurement concentration range, and this phenomenon can be resolved. On the other hand, when the object to be measured is an antibody, there is no particular limitation, but examples include substances used in clinical tests, such as human immunoglobulin, antithyroid peroxidase antibody, antithyroglobulin antibody, and TSH receptor antibody.

[0009] In the present invention, the solid-phase antibody refers to an antibody against the antigen to be measured, which is immobilized on a solid-phase carrier such as beads or microparticles, and is insoluble in the solution in which the antigen-antibody reaction is carried out. However, the antibody does not have to be directly bound to the solid phase; for example, it may be bound via an avidin-biotin bond. Specifically, it also includes a soluble antibody bound to biotin for binding to an avidin-solid phase complex.

[0010] In the present invention, the solid-phase antigen refers to an antigen that is immobilized on a solid-phase carrier such as beads or microparticles and that is insoluble in the solution in which the antigen-antibody reaction is carried out. However, the antigen does not have to be directly bound to the solid phase; for example, it may be bound via an avidin-biotin bond. Specifically, it also includes soluble antigens bound to biotin for binding to an avidin-solid-phase complex.

[0011] The labeled antibody refers to an antibody against the antigen or antibody to be measured, to which a label is bound, and which is soluble in the solution in which the antigen-antibody reaction is carried out. However, the label does not have to be bound directly to the antibody; for example, the label may be bound via an avidin-biotin bond. Specifically, the labeled antibody also includes a soluble antibody bound to biotin for binding to an avidin-label complex.

[0012] The labeled antigen refers to an antigen to be measured, which is bound to a label, and is soluble in a solution in which an antigen-antibody reaction is carried out. However, the label does not have to be bound directly to the antigen; for example, the label may be bound via an avidin-biotin bond. Specifically, the labeled antigen includes soluble antigens bound to biotin for binding to an avidin-label complex.

[0013] As the solid phase, beads, microparticles, or wells can be used. Microparticles are particularly preferred, and may be inorganic materials such as glass, metal, or ceramics, or organic materials such as high molecular weight polymers. These beads or microparticles may also contain magnetic materials. The particle diameter of the microparticles is preferably 0.1 to 50 μm, and more preferably 1 to 10 μm. The particle diameter of the beads is preferably 0.1 to 5 mm, and more preferably 0.5 to 2 mm.

[0014] The label is not particularly limited, but examples thereof include enzymes, radioisotopes, etc., and enzymes are particularly preferred, such as alkaline phosphatase and peroxidase.

[0015] When an enzyme is used as a label, a substrate that reacts with the enzyme can be used, such as a fluorescent substrate, a chemiluminescent substrate, a bioluminescent substrate, or a chromogenic substrate.

[0016] Immunoassays using these methods are solid-phase methods, and include sandwich and competitive assays. Furthermore, the antibody or antigen does not have to be directly bound to the solid phase, but rather is bound to the solid phase in a reaction step subsequent to the antigen-antibody reaction, such as an immunoassay using a biotinylated antigen in a sandwich assay followed by an avidin-solid phase complex reaction. Furthermore, the antibody does not have to be directly bound to the label, but rather is bound to the label in a reaction step subsequent to the antigen-antibody reaction, such as an immunoassay using a biotinylated antibody in a sandwich assay followed by an avidin-label complex reaction.

[0017] The antibody in the present invention may be a polyclonal antibody or a monoclonal antibody, and may be derived from any animal species that produces antibodies, such as rabbits, goats, sheep, pigs, horses, mice, or rats. The antibody may be in the form of a complete antibody or an F(ab') fragment obtained by cleaving the complete antibody by enzymatic or chemical treatment. ２ The antibody may be an antibody fragment such as Fab' or Fab'. Furthermore, scFv (single-chain variable region fragment) or VHH antibody may also be used.

[0018] The antigen that competes with the antigen to be measured is not particularly limited as long as it is an antigen that competes for binding to an antibody that binds to the antigen to be measured, and may be the same antigen as the antigen to be measured or a different antigen. When the competing antigen is an antigen that is different from the antigen to be measured, it is preferably an antigen that has the same epitope as the epitope recognized by the antibody that binds to the antigen to be measured.

[0019] The antifoaming agent used in the present invention is not particularly limited as long as it is an antifoaming agent that can eliminate bubbles generated during dissolution while maintaining the three-dimensional structure of the antibody or antigen. For example, an emulsion-type water-soluble silicone-based antifoaming agent, a self-emulsifying water-soluble silicone-based antifoaming agent, etc. can be used. Preferred examples of the emulsion-type water-soluble silicone-based antifoaming agent include KM-7750 (Shin-Etsu Chemical Co., Ltd.), KM-71 (Shin-Etsu Chemical Co., Ltd.), KM-75 (Shin-Etsu Chemical Co., Ltd.), and Antifoam SI (Wako Pure Chemical Industries, Ltd.), and preferred examples of the self-emulsifying water-soluble silicone-based antifoaming agent include KS-538 (Shin-Etsu Chemical Co., Ltd.). Among these, KM-7750 (Shin-Etsu Chemical Co., Ltd.) and Antifoam SI (Wako Pure Chemical Industries, Ltd.) are more preferred. Furthermore, one type of antifoaming agent may be added, or two or more types may be added. For example, only an emulsion-type water-soluble silicone-based antifoaming agent may be used, only a self-emulsifying water-soluble silicone-based antifoaming agent may be used, or both may be combined. The lower limit of the concentration range of the antifoaming agent in the solution before freeze-drying is preferably 0.0001% or more, more preferably 0.0005% or more, and even more preferably 0.001% or more (volume / volume), and the upper limit is preferably 0.1% or less, more preferably 0.05% or less, and even more preferably 0.01% or less (volume / volume). Specifically, the antifoaming agent is preferably in a concentration range of 0.0001 to 0.1% (volume / volume) in the solution before freeze-drying, more preferably 0.0005 to 0.05% (volume / volume), and particularly preferably 0.001 to 0.01% (volume / volume), for example, 0.005% (volume / volume) is exemplified.

[0020] Examples of the solution used in this process include buffer solutions and water, as well as solutions containing sugars, salts, proteins, and surfactants. Examples of sugars that can be used include sucrose, mannitol, trehalose, and inositol. Examples of proteins that can be used include bovine serum albumin, collagen peptides, and skim milk. Examples of surfactants that can be used include anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants. Examples of buffer solutions that can be used include Tris, MOPSO, MOPS, and MES, and examples of salts that can be used include sodium chloride, potassium chloride, magnesium chloride, and zinc chloride. In addition to these, other reagent components can also be present during freeze-drying, if necessary.

[0021] In the present invention, it is essential that the freeze-dried reagent contains an antifoaming agent coexisting with a solid-phase antibody or solid-phase antigen, but it may also contain a freeze-dried reagent containing an antifoaming agent coexisting with a labeled antibody or labeled antigen.

[0022] When producing the freeze-dried reagent of the present invention, the solid-phase antibody or solid-phase antigen in the solution may be frozen and freeze-dried after precipitation. However, taking into consideration dispersibility after redissolution, it is preferable to freeze and freeze-dry the suspension of the solid-phase antibody or solid-phase antigen immediately after dispersion, i.e., in the state in which the solid-phase antibody or solid-phase antigen is dispersed.

[0023] In this manner, the freeze-dried immunoreaction reagent of the present invention can be produced.

[0024] As described above, the kit of the present invention comprises a freeze-dried reagent in which an antifoaming agent is coexisted with a solid-phase antibody or solid-phase antigen, and a reagent containing a labeled antibody or labeled antigen. In this case, the reagent containing the labeled antibody or labeled antigen may further contain an antifoaming agent or may be freeze-dried.

[0025] According to the present invention, it is possible to obtain a freeze-dried immunoreaction reagent and a kit thereof which have good measurement reproducibility and are used for in vitro diagnostics, etc. For example, the immunoassay reagent and kit thereof obtained according to the present invention enable accurate measurements, and therefore high-precision measurements can also be achieved by the present invention.

[0026] FIG. 1 shows the measurement results of the freeze-dried reagent prepared in Example 1. FIG. 2 shows the measurement results of the freeze-dried reagent prepared in Comparative Example 1. FIG. 3 shows the measurement results of the freeze-dried reagent prepared in Example 2. FIG. 4 shows the measurement results of the freeze-dried reagent prepared in Comparative Example 2. FIG. 5 shows the state of the freeze-dried reagents prepared in Example 1 and Comparative Example 1 immediately after dissolution. FIG. 6 shows the state of the freeze-dried reagents prepared in Example 1 and Comparative Example 1 35 seconds after dissolution. FIG. 7 shows the measurement results of the freeze-dried reagent prepared in Example 4. FIG. 8 shows the measurement results of the freeze-dried reagent prepared in Comparative Example 3.

[0027] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0028] Each measurement was performed by the one-step delay method or the one-step competitive method using an automatic chemiluminescent enzyme immunoassay device (AIA-CL2400, manufactured by Tosoh Corporation) as the immunoassay device and immunoreaction reagents for the device as the immunoassay reagents. Each immunoreaction reagent was prepared as described below.

[0029] Example 1 Anti-rabbit-goat antibody-immobilized magnetic particles were added to a Tris buffer solution (placed in one of the two-hole containers shown in FIG. 5 ) containing anti-aldosterone-rabbit antibody, trehalose, mannitol, potassium chloride, magnesium chloride, zinc chloride, sodium heparin, sodium caseinate, Proclin 950, bovine serum albumin, and collagen peptide, and a silicone emulsion-type antifoaming agent (KM-7750 (Shin-Etsu Chemical Co., Ltd.)) was further added to a concentration of 0.005% (volume / volume). This suspension was stirred to completely disperse the magnetic particles, and immediately thereafter frozen, followed by freeze-drying. Next, alkaline phosphatase-labeled aldosterone was added to a Tris buffer solution containing trehalose, collagen peptide, magnesium chloride, zinc chloride, and Proclin 300 (which was placed in the other well of the two-well container), and a silicone emulsion-type antifoaming agent (KM-7750 (Shin-Etsu Chemical Co., Ltd.)) was added to a concentration of 0.005% (volume / volume), followed by freeze-drying.

[0030] Next, using the reagent prepared in this manner, EDTA plasma samples were measured using the automated immunoassay device by the one-step delay method, and the luminescence intensity of the chemiluminescent substrate, which is a substrate for alkaline phosphatase, was measured. The EDTA plasma samples were measured 30 times, and the average value was used as the measured value. Furthermore, the measurement reproducibility was calculated based on the measured values. The average luminescence intensity was 32,392 (RLU / sec.), and the reproducibility (CV: Coefficient of Variation) was good at 1.4%. The results are shown in Table 1 and Figure 1.

[0031]

[0032] Comparative Example 1: Anti-rabbit-goat antibody-immobilized magnetic microparticles were added to a Tris buffer solution (one of the two-well containers shown in FIG. 5 ) containing anti-aldosterone-rabbit antibodies, trehalose, mannitol, potassium chloride, magnesium chloride, zinc chloride, sodium heparin, sodium caseinate, Proclin 950, bovine serum albumin, and collagen peptide. The suspension was stirred to completely disperse the magnetic microparticles, immediately frozen, and then lyophilized (no antifoaming agent added). Next, alkaline phosphatase-labeled aldosterone was added to a Tris buffer solution (the other of the two-well containers) containing trehalose, collagen peptide, magnesium chloride, zinc chloride, and Proclin 300. A silicone emulsion-type antifoaming agent (KM-7750 (Shin-Etsu Chemical Co., Ltd.)) was added to a concentration of 0.005% (volume / volume), and the resulting solution was lyophilized.

[0033] Next, using the reagent thus prepared, a test was carried out in the same manner as in Example 1. The average luminescence intensity was 32,224 (RLU / sec), and the reproducibility was 4.1%, which was inferior to that of Example 1. The results are shown in Table 1 and Figure 2.

[0034] Example 2: Anti-FITC mouse monoclonal antibody-immobilized magnetic particles were added to a Tris buffer solution (placed in one well of the two-well container shown in Figure 5) containing an FITC-labeled S-adenosyl-L-homocysteine ​​BSA conjugate, trehalose, mannitol, dextran, magnesium chloride, zinc chloride, Proclin 950, gentamicin sulfate, collagen peptide, dissociation inhibitor HBR-1, and ultra-low activity alkaline phosphatase. This suspension was stirred to completely disperse the magnetic particles, and immediately thereafter frozen, followed by lyophilization. Furthermore, alkaline phosphatase-labeled anti-S-adenosyl-L-homocysteine ​​mouse monoclonal antibody was added to a Tris buffer solution containing trehalose, mannitol, magnesium chloride, zinc chloride, Proclin 950, gentamicin sulfate, and collagen peptide, and a silicone emulsion-type antifoaming agent (KM-7750 (Shin-Etsu Chemical Co., Ltd.)) was further added to a concentration of 0.01% (volume / volume). The mixture was placed in the same well of the two-well container and freeze-dried.

[0035] Next, using the reagent prepared in this manner, human serum samples were measured by the one-step competitive assay using the automated immunoassay device, and the luminescence intensity of the chemiluminescent substrate, which is a substrate for alkaline phosphatase, was measured. The human serum samples were measured 60 times, and the average value was taken as the measured value. Furthermore, the measurement reproducibility was calculated based on the measured values. The average luminescence intensity was 7074 (RLU / sec.), and the reproducibility (CV: Coefficient of Variation) was 4.2%, which was good. The results are shown in Table 2 and Figure 3.

[0036]

[0037] Comparative Example 2: Anti-FITC mouse monoclonal antibody-immobilized magnetic microparticles were added to a Tris buffer solution (placed in one of the wells of the two-well container shown in Figure 5) containing an FITC-labeled S-adenosyl-L-homocysteine ​​BSA conjugate, trehalose, mannitol, dextran, magnesium chloride, zinc chloride, Proclin 950, gentamicin sulfate, collagen peptide, dissociation inhibitor HBR-1, and ultra-low activity alkaline phosphatase. This suspension was stirred to completely disperse the magnetic microparticles, immediately frozen, and then lyophilized. Furthermore, alkaline phosphatase-labeled anti-S-adenosyl-L-homocysteine ​​mouse monoclonal antibody was added to a Tris buffer solution containing trehalose, mannitol, magnesium chloride, zinc chloride, Proclin 950, gentamicin sulfate, and collagen peptide, and the resulting solution was placed in the same well of the two-well container and lyophilized (without the addition of an antifoaming agent).

[0038] Next, using the reagent prepared in this manner, a test was carried out in the same manner as in Example 2. The average luminescence intensity was 7214 (RLU / sec), and the reproducibility was 14.0%, which was inferior to that of Example 2. The results are shown in Table 2 and Figure 4.

[0039] Example 3 In two-well containers containing the freeze-dried reagents prepared in Example 1 and Comparative Example 1, a dissolving solution was added to the well containing the magnetic microparticles to dissolve the reagent, and the occurrence of bubbles was confirmed. The freeze-dried reagent prepared in Example 1 disappeared 35 seconds after dissolution, but the freeze-dried reagent prepared in Comparative Example 1 did not disappear even after 1 minute had passed, and bubbles remained. Figure 5 shows the state immediately after dissolution, and Figure 6 shows the state 35 seconds after dissolution. In Figures 5 and 6, arrows indicate the areas where bubbles exist.

[0040] In the case of Comparative Example 1, which did not contain an antifoaming agent, the bubbles that were generated when the freeze-dried reagent was dissolved did not disappear, and the reproducibility of 30 measurements was 4.1%. In contrast, in the case of Example 1, in which an antifoaming agent was added, the bubbles that were generated when the freeze-dried reagent was dissolved disappeared, and the reproducibility of 30 measurements was significantly improved to 1.4%. This shows that the addition of an antifoaming agent suppressed the variation caused by bubbles when the freeze-dried reagent was dissolved, and improved the reproducibility of the immune reaction reagent.

[0041] Example 4 A freeze-dried reagent was prepared in the same manner as in Example 1, except that a silicone emulsion type antifoaming agent, Antifoam SI (Wako Pure Chemical Industries, Ltd.), was used as the antifoaming agent.

[0042] Next, using the reagent prepared in this manner, 30 measurements were performed in the same manner as in Example 1, and the average value was taken as the measured value. Furthermore, the measurement reproducibility was calculated based on the measured values. The average value of the luminescence intensity was 24,115 (RLU / sec.), and the reproducibility (CV: Coefficient of Variation) was 1.8%, which was good. The results are shown in Table 3 and Figure 7.

[0043]

[0044] Comparative Example 3 A freeze-dried reagent was prepared in the same manner as in Comparative Example 1, except that a silicone emulsion type antifoaming agent, Antifoam SI (Wako Pure Chemical Industries, Ltd.), was used as the antifoaming agent.

[0045] Next, using the reagent thus prepared, a test was carried out in the same manner as in Example 4. The average luminescence intensity was 23,022 (RLU / sec), and the reproducibility was 4.1%, which was inferior to that of Example 4. The results are shown in Table 3 and Figure 8.

Claims

1. An immune reaction reagent comprising a freeze-dried reagent containing a solid-phase antibody against an antigen to be measured and an antifoaming agent.

2. An immune reaction reagent comprising a freeze-dried reagent containing a solid-phase antigen for an antibody to be measured and an antifoaming agent.

3. An immune reaction reagent comprising: a freeze-dried reagent containing a solid-phase antigen that competes with the antigen to be measured for binding to an antibody against the antigen, and an antifoaming agent.

4. An immune reaction reagent according to any one of claims 1 to 3, wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent.

5. A method for producing an immune reaction reagent, characterized by coexisting a solid-phase antibody against the antigen to be measured and an antifoaming agent in a solution and freeze-drying the solution.

6. A method for producing an immune reaction reagent, characterized by coexisting a solid-phase antigen against the antibody to be measured and an antifoaming agent in a solution and freeze-drying the solution.

7. A method for producing an immune reaction reagent, characterized in that a solid-phase antigen that competes with the antigen to be measured for binding to an antibody against the antigen to be measured and an antifoaming agent are coexisted in a solution and freeze-dried.

8. A method for producing an immune reaction reagent according to any one of claims 5 to 7, wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent.

9. A method for producing an immune reaction reagent according to any one of claims 5 to 7, wherein the solid-phase antibody or the solid-phase antigen is dispersed as a suspension and immediately thereafter frozen and lyophilized.

10. An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antibody against an antigen to be measured and an antifoaming agent; and a reagent containing a labeled antibody capable of binding to the antigen.

11. An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antibody against an antigen to be measured and an antifoaming agent; and a reagent containing a labeled antigen capable of binding to the solid-phase antibody.

12. An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antigen for an antibody to be measured and an antifoaming agent; and a reagent containing a labeled antibody capable of binding to the antigen.

13. An immune reaction reagent kit comprising: a freeze-dried reagent containing a solid-phase antigen that competes with the antigen to be measured for binding to an antibody against the antigen, and an antifoaming agent; and a reagent containing a labeled antibody capable of binding to the solid-phase antigen and the antigen to be measured.

14. An immune reaction reagent kit according to any one of claims 10 to 13, wherein the antifoaming agent is an emulsion-type water-soluble silicone-based antifoaming agent and / or a self-emulsifying water-soluble silicone-based antifoaming agent.

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