Immunodiagnostic reagent accelerant and preparation method thereof

Abstract

The invention provides an immunodiagnostic reagent accelerant and a preparation method thereof. Comprising a reinforcing agent, a pH regulator, protective protein, a surfactant, inorganic salt and a bacteriostatic agent. The immunodiagnostic reagent accelerant is used as a matched enhancing reagent of a common immunodiagnostic reagent, the binding efficiency of an antigen and an antibody in the immunodiagnostic reagent can be effectively improved, and a detection signal is increased by improving the binding efficiency, so that the improvement of the detection sensitivity and the reduction of the detection time are realized; the detection efficiency is improved.

Description

Technical Field

[0001] This invention relates to the field of immunodiagnostic technology, and in particular to an immunodiagnostic reagent enhancer and its preparation method. Background Technology

[0002] In today's in vitro diagnostics field, immunodiagnostics is the most diverse, widely applied, and largest-scale technological area. Specifically, it involves methods for capturing, enriching, and detecting antigens, haptens, and antibodies based on the specific binding between them. Mainstream immunodiagnostic methods include enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay, immunochromatography, immunoturbidimetry, and flow cytometry. Immunodiagnostic technology has long focused on improving sensitivity, specificity, and detection efficiency, with the key being enhancing the efficiency and rate of antigen-antibody binding.

[0003] Antigens and antibodies bind together using non-bonded forces such as dispersion forces, electrostatic forces, and hydrogen bonds. Dispersion forces and electrostatic forces play a crucial role in bringing the antigens and antibodies closer together, often in the early stages of binding. These forces shorten the distance between them and adjust their relative positions. When they reach their optimal positions, hydrogen bonds form, ensuring a strong bond. However, antigens and antibodies are often proteins with numerous hydrophilic polar groups on their surfaces, such as -COOH, -NH2, and -OH. These groups form hydrogen bonds with water molecules, creating a hydration layer of approximately 1-1.5 nm thick, essentially "fixed water molecules." Additionally, different ions in the buffer solution are dispersed on the protein surface through electrostatic adsorption. The presence of this hydration layer separates protein particles and generates electrostatic repulsion, preventing direct protein aggregation due to collisions and hindering the interaction between antigens and antibodies, thus reducing the binding efficiency. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention relates to an enhancer for immunodiagnostic reagents. Specifically, adding an enhancer to commercially available immunodiagnostic reagents can disrupt the hydration layer on the protein surface, effectively enhancing the binding efficiency of antigens and antibodies during detection, improving reagent performance, and without compromising protein stability. The immunodiagnostic enhancer described in this invention can be used in common immunodiagnostic fields such as enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay, immunochromatography, and immunoturbidimetry.

[0005] In one aspect, the present invention provides an immunodiagnostic enhancer, comprising an enhancer, a pH adjuster, a protective protein, a surfactant, an inorganic salt, and an antibacterial agent.

[0006] According to embodiments of the present invention, the reinforcing agent may be a monool with a carbon chain length of 1-20, such as propanol, butanol, hexanol, etc.; a polyol with a carbon chain length of 1-2, such as propylene glycol, hexanediol, tetraethylene glycol, etc.; a polymer or copolymer of polyethylene glycol (PEG, PEO), polypropylene glycol (PPG, PPO), polytetrahydrofuran (PTHF), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polymethacrylic acid (PMA), linear polyacrylamide (LPAM), etc., with a degree of polymerization of 10-200,000; or one or a mixture of multiple monosaccharides or polysaccharides such as glucose, fructose, sucrose, dextran, glycogen, etc.

[0007] According to an embodiment of the present invention, the pH adjuster may be one or more of the following: disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, sodium citrate, 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), 2-morpholine ethanesulfonic acid (MES), 3-morpholine propanesulfonic acid (MPOS), 2-[[tris(hydroxymethyl)methyl]amino]ethanesulfonic acid (TES), piperazine-N,N'-di(2-ethanesulfonic acid) (PIPES), tris(hydroxymethyl)aminomethane (Tris), tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl), and tris(hydroxymethyl)aminopropanesulfonic acid (TAPS).

[0008] According to an embodiment of the present invention, the protective protein can be a purified protein, polypeptide, or oligopeptide. The protein source can be obtained through processes such as purification of natural substances, chemical synthesis, polypeptide synthesis, or recombinant protein. Specifically, it can be one or more of the following protective proteins: human serum albumin (hereinafter referred to as HSA), bovine serum albumin (hereinafter referred to as BSA), ovalbumin (hereinafter referred to as OVA), gelatin, casein and its derivatives, such as hydrolyzed casein, oligopeptides, polypeptides, and fish serum albumin. The preferred protective proteins are HSA, BSA, OVA, casein and its derivatives, and fish serum albumin. More preferably, it is HSA, BSA, and fish serum albumin.

[0009] According to embodiments of the present invention, the surfactant can be one or more of nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants. Specifically, nonionic surfactants include polysorbates and alkylphenol polyoxyethylene ethers, such as Tween 20, Tween 40, Tween 60, Tween 80, and Triton, with Tween 20 and Triton being preferred; anionic surfactants include sulfonate surfactants, with sodium dodecyl sulfonate (hereinafter referred to as SDS) being preferred; cationic surfactants include quaternary ammonium salt surfactants, including dodecyltrimethylammonium bromide (DTAB), tetradecyltrimethylammonium bromide, and hexadecyltrimethylammonium bromide (CTAB); and amphoteric surfactants include Tetronic 1307 (hereinafter referred to as S9), more preferably Tween 20, Tween 40, and S9.

[0010] According to an embodiment of the present invention, the inorganic salt may be one or a mixture of common inorganic salts such as sodium chloride, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, sodium citrate, sodium ascorbate, potassium phosphate, sodium phosphate, ammonium sulfate, ammonium chloride, and sodium gluconate.

[0011] According to embodiments of the present invention, the antibacterial agent can be an inorganic or organic compound with antibacterial activity, specifically sodium azide, potassium sorbate, potassium sorbitol, sodium phenylpropionate, isothiazolinone, methylchloroisothiazolinone, methylisothiazolinone, Proclin-300, etc., with preferred antibacterial agents being sodium azide, methylchloroisothiazolinone, methylisothiazolinone, and Proclin-300, and more preferably sodium azide and Proclin-300.

[0012] According to an embodiment of the present invention, the immunodiagnostic enhancer comprises any one of the following: (1) PB, NaCl, BSA, Tween 20, butanol, sodium azide; (2) MES, NaCl, BSA, Tween 20, butanol, Proclin 300; (3) MES, NaCl, ammonium sulfate, BSA, S9, PEG10000, Proclin 300; (4) PB, NaCl, ammonium sulfate, BSA, Tween 40, PEG20000, Proclin 300; (5) Tris, NaCl, ammonium sulfate, HSA, Tween 40, PEG10000, sodium azide. Preferably, the immunodiagnostic enhancer comprises any one of the following: (1) PB, NaCl, BSA, Tween 20, butanol, sodium azide; (2) MES, NaCl, BSA, Tween 20, butanol, Proclin 300.

[0013] According to an embodiment of the present invention, the content of the enhancer is 0.01-30%, preferably 5-15%, and more preferably 10%. The content of the pH adjuster is 1-1000 mM, preferably 10-200 mM, and more preferably 50 mM. The content of the protective protein is 0.01-20%, preferably 0.5-5%, and more preferably 1%. The content of the surfactant is 0.01-10%, preferably 0.05-2%, and more preferably 0.1%. The content of the inorganic salt is 50 mM-1 M, preferably 150 mM. The content of the antibacterial agent is 0.005-5%, preferably 0.05%.

[0014] According to a preferred embodiment of the present invention, the immunodiagnostic enhancer comprises 1-1000mM MES at pH 6.0, 50mM-1M NaCl, 0.01-20% BSA, 0.01-10% Tween 20, 0.01-30% butanol, and 0.005-5% Proclin 300.

[0015] According to an embodiment of the present invention, the immunodiagnostic enhancer can significantly improve the specific binding efficiency of antigens and antibodies.

[0016] According to an embodiment of the present invention, compared with a control test without the addition of the immunodiagnostic enhancer, the detection signal intensity (such as OD value, relative luminescence value, or fluorescence intensity) can be increased by at least 100% after using the immunodiagnostic enhancer.

[0017] In a second aspect, the present invention provides a method for preparing the immunodiagnostic enhancer, comprising the step of mixing an enhancer, a pH adjuster, a protective protein, a surfactant, an inorganic salt, and an antibacterial agent.

[0018] In a third aspect, the present invention provides a method for improving the efficiency of an immunodiagnostic reaction, comprising the step of adding the immunodiagnostic enhancer to an immunodiagnostic reaction system, wherein the immunodiagnostic reaction is selected from enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay, or immunochromatographic reaction.

[0019] According to an embodiment of the present invention, the proportion of the immunodiagnostic enhancer added to the immunodiagnostic reaction system is 10%.

[0020] According to an embodiment of the present invention, in the enzyme-linked immunosorbent assay (ELISA), the immunodiagnostic enhancer is added to a 96-well plate, followed by the addition of a sample, enzyme conjugate, and substrate for incubation and color development; in the chemiluminescence reaction, the immunodiagnostic enhancer replaces the solvent of the magnetic bead components; and in the immunochromatographic reaction, the immunodiagnostic enhancer is added to the sample buffer to form a mixed dilution.

[0021] According to an embodiment of the present invention, after using the immunodiagnostic enhancer, the detection signal intensity is increased by at least twice compared to the control group without the enhancer.

[0022] In a fourth aspect, the present invention provides the use of the aforementioned immunodiagnostic enhancer in the preparation of reagents that improve the efficiency of immunodiagnostic reactions, wherein the immunodiagnostic reaction is selected from enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay, or immunochromatographic reaction.

[0023] According to an embodiment of the present invention, the improvement in the efficiency of the immunodiagnostic reaction is manifested in improving the antigen-antibody binding efficiency and / or increasing the intensity of the detection signal, including OD value, relative luminescence value or fluorescence intensity.

[0024] In a fifth aspect, the present invention provides an immunodiagnostic kit, comprising the aforementioned immunodiagnostic enhancer, and one or more of enzyme-linked immunosorbent assay (ELISA) reagents, chemiluminescent reagents, or immunochromatographic reagents.

[0025] According to an embodiment of the present invention, the kit further includes a sample diluent, a magnetic bead component, an enzyme conjugate, a substrate, or a sample buffer, wherein the immunodiagnostic stimulant is used in combination with the sample diluent, the magnetic bead component, or the sample buffer.

[0026] Compared with the prior art, the present invention has the following beneficial effects: The immunodiagnostic reagent enhancer described in this invention is a common immunodiagnostic reagent that is used as a complementary enhancer. It can effectively increase the binding efficiency of antigens and antibodies in immunodiagnostic reagents. By increasing the binding efficiency, the detection signal is increased, thereby improving detection sensitivity and reducing detection time (Examples 2-4), thus improving detection efficiency. Detailed Implementation

[0027] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0028] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0029] Terminology Explanation Immunodiagnostic enhancers are chemical reagent combinations used to improve the efficiency of immunodiagnostic reactions (such as enzyme-linked immunosorbent assay (ELISA), chemiluminescent immunoassay (CLIA), or immunochromatographic assay (LFA). They enhance signal intensity (such as OD value, luminescence value, or fluorescence intensity) and reaction rate through mechanisms such as optimizing antigen-antibody binding, reducing surface tension, stabilizing proteins, and adjusting pH. In this invention, it comprises enhancers, pH adjusters, protective proteins, surfactants, inorganic salts, and antibacterial agents, typically added at a ratio of 10%, which can at least double the signal intensity, making it suitable for medical testing, food safety, and other fields.

[0030] Enhancers: Additives used to enhance immune response signals and efficiency. These include: Monohydric alcohols: Alcohols containing one hydroxyl group (-OH) in their molecule (e.g., propanol, butanol, hexanol). Polyhydric alcohols: Alcohols containing multiple hydroxyl groups (e.g., propylene glycol, hexanediol, tetraethylene glycol). High molecular weight polymers (e.g., PEG, PVP, PVA): Hydrophilic polymers with a degree of polymerization of 10-200,000. Sugars (e.g., glucose, sucrose, dextran): Monosaccharides or polysaccharides. Concentration: 0.01-30% (preferably 10%). Overall, enhancers improve antigen-antibody binding efficiency and reduce false positives.

[0031] pH adjusters: Compounds used to maintain the stability of solution pH, such as phosphates (Na2HPO4 / NaH2PO4), MES, Tris-HCl, etc. pH affects protein charge and binding affinity. In this invention, the concentration is 1-1000 mM (preferably 50 mM), and the pH range is typically 6-8 to optimize enzyme activity or antibody binding.

[0032] Protective proteins: Proteins used to protect antibodies or antigens from degradation or nonspecific adsorption, such as HSA (human serum albumin), BSA (bovine serum albumin), OVA (ovalbumin), casein, or fish serum albumin. Sources include natural extracts or recombinant expression, at concentrations of 0.01-20% (preferably 1%). In this context, they act as blocking agents, reducing background noise and improving the signal-to-noise ratio.

[0033] Surfactants: Compounds used to reduce the surface tension of solutions, including: nonionic surfactants (such as Tween 20, Tween 40, Triton X-100); anionic surfactants (such as SDS / sodium dodecyl sulfate); cationic surfactants (such as CTAB): positively charged, rarely used; amphoteric surfactants (such as S9 / Tetronic 1307): charge varies with pH. Concentration: 0.01-10% (preferably 0.1%), with Tween 20 / S9 preferred in this invention.

[0034] Inorganic salts, such as NaCl, KCl, and (NH4)2SO4 (ammonium sulfate), at concentrations of 50 mM to 1 M (preferably 150 mM), regulate ionic strength, affecting protein solubility and binding affinity. In immune responses, they can promote salt bridge formation or precipitate impurities, thereby increasing specificity.

[0035] Antimicrobial agents: Compounds that prevent microbial contamination, such as sodium azide (NaN3) and Proclin-300. Concentration: 0.005-5% (preferably 0.05%). Sodium azide inhibits respiratory enzymes, while Proclin-300 is a broad-spectrum bactericide used to extend the shelf life of reagents.

[0036] Immunodiagnostic reactions: Detection methods based on antigen-antibody specific binding, including ELISA (enzyme-linked immunosorbent assay, using enzyme markers to generate color signals), chemiluminescence (using luminescent substrates to generate light signals), and immunochromatography (lateral flow assay, using markers such as colloidal gold to generate visible lines). In this invention, the promoter enhances binding efficiency, at least doubling the signal.

[0037] Signal intensity detection: an indicator of the quantification of reaction results, such as OD value (absorbance, in ELISA), relative luminescence value (RLU, in chemiluminescence), or fluorescence intensity (in immunochromatography). This invention improves upon this by at least twofold.

[0038] % indicates the mass-volume ratio, such as g / 100mL.

[0039] Example 1: Preparation of Accelerators with Different Formulations This embodiment describes the preparation of different formulations of stimulants. Different formulations of immunodiagnostic stimulants were prepared according to the formulations in Table 1.

[0040] Table 1: List of different accelerator formulations

[0041] Example 2: Use of Immunodiagnostic Enhancers in Enzyme-Linked Immunosorbent Assay Reagents This embodiment uses commercially available enzyme-linked immunosorbent assay (ELISA) reagents. Two control groups were established using no reagents and the same amount of purified water, respectively, to test the five different formulations of accelerators obtained in Example 1. The specific implementation method is as follows: ① Add 10 μL of control or accelerator to the 96-well plate in advance; ② Follow the instructions: add the sample and incubate, wash the plate, add the enzyme conjugate and incubate again, wash again, add the substrate and incubate, stop the incubation after color development and read the corresponding OD value.

[0042] Table 2: Use of Immunodiagnostic Enhancers in Enzyme-Linked Immunosorbent Assays

[0043] The results above show that the OD values ​​detected in the experimental groups using promoters were higher than those in the control group. In particular, the results of promoters 1 and 2 were twice as high as those of the control group. This means that in the experiments using promoters, the promoters can make the binding efficiency of antigen and antibody higher.

[0044] Example 3: Application of Immunodiagnostic Enhancers in Chemiluminescent Reagents This embodiment uses commercially available chemiluminescence reagents. Two control groups were established by adding no reagents and adding an equal amount of purified water. The accelerators of the five different formulations obtained in Example 1 were tested. The specific implementation method is as follows: In the existing reagent components, 10% of the solvent was replaced in the magnetic bead components for the control group and the accelerator experimental group, respectively. Specifically, 5 mL of magnetic bead working solution was taken, and after magnetic separation, 0.5 mL of supernatant was taken. Then, 0.5 mL of the control group and the experimental group were added respectively for subsequent normal testing. The test samples were high-value samples with serial dilution.

[0045] Table 3: Use of Immunodiagnostic Enhancers in Chemiluminescence Reactions

[0046] The results above are consistent with those of enzyme-linked immunosorbent assay (ELISA). In the experimental groups using promoters, the relative luminescence values ​​were higher than those in the control group. Particularly with promoters 1 and 2, the S1 and S2 values ​​were twice that of the control group, indicating that the promoters significantly increased the antigen-antibody binding efficiency. Furthermore, compared to the control group, the addition of promoters improved the signal-to-noise ratio (S1 / S0), demonstrating that the promoters effectively increased the detection sensitivity of the reagent.

[0047] Example 4: Application of Immunodiagnostic Enhancers in Immunochromatographic Reagents This embodiment uses commercially available immunochromatographic reagents. Two control groups were established using no reagents and an equal amount of purified water, respectively, to test the five different formulations of accelerators obtained in Example 1. The specific implementation method is as follows: Add an additional 10% solvent to the sample buffer in the kit to form a mixed sample dilution. Specifically, take 1 mL of sample buffer and add 0.1 mL of control and experimental groups for subsequent normal testing. The test sample is a high-value sample with a gradient dilution.

[0048] Table 4: Use of Immunodiagnostic Enhancers in Fluorescent Immunochromatographic Reactions

[0049] The results above are consistent with those of enzyme-linked immunosorbent assay (ELISA). In the experimental groups using promoters, the detected fluorescence intensity values ​​were higher than those in the control group. Particularly in the experiments using promoters 1 and 2, the S1 and S2 values ​​were twice that of the control group, indicating that the promoters significantly increased the antigen-antibody binding efficiency. Furthermore, compared to the control group, the addition of promoters improved the signal-to-noise ratio (S1 / S0), demonstrating that the promoters effectively increased the detection sensitivity of the reagent.

[0050] Example 5: Synergistic effect of immunodiagnostic enhancers This embodiment uses commercially available chemiluminescence reagents. Two control groups were established using no reagents and the same amount of purified water. The five different formulations of the accelerators shown in the table below were tested. The specific implementation method is as follows: In the existing reagent components, 10% of the solvent was replaced in the magnetic bead components for the control group and the accelerator experimental group, respectively. Specifically, 5 mL of magnetic bead working solution was taken, and after magnetic separation, 0.5 mL of supernatant was taken. Then, 0.5 mL of the control group and the experimental group were added respectively for subsequent normal testing. The test samples were S0 and S1 samples, and the signal-to-noise ratio of S1 / S0 was calculated.

[0051] Table 5: Synergistic effect of immunodiagnostic enhancers

[0052] The results show that the signal-to-noise ratio (SNR) S1 / S0 improved to varying degrees with the addition of different accelerators. Specifically, adding only butanol as an enhancer improved the SNR compared to the control group, and the SNR decreased as the proportion of butanol decreased. Adding the antibacterial agent Proclin 300 did not significantly improve the SNR, but adding Tween 20 significantly improved it. Removing the enhancer butanol (accelerator-10) resulted in a decrease in the measured SNR. Finally, accelerator-2 exhibited the best SNR, indicating that all components in the accelerator system showed a synergistic effect, especially Tween 20 and butanol, which had a synergistic effect on improving the SNR. Furthermore, the improvement in SNR varied with different formulations.

[0053] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. An immunodiagnostic enhancer, characterized in that, This includes enhancers, pH adjusters, protective proteins, surfactants, inorganic salts, and antibacterial agents.

2. The immunodiagnostic enhancer as described in claim 1, characterized in that, The reinforcing agent can be a single alcohol with a carbon chain length of 1-20, such as propanol, butanol, and hexanol; a polyol with a carbon chain length of 1-2, such as propylene glycol, hexanediol, and tetraethylene glycol; a high molecular weight polymer or copolymer with a degree of polymerization of 10-200,000, such as polyethylene glycol (PEG, PEO), polypropylene glycol (PPG, PPO), polytetrahydrofuran (PTHF), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polymethacrylic acid (PMA), and linear polyacrylamide (LPAM); or one or more monosaccharides or polysaccharides such as glucose, fructose, sucrose, dextran, and glycogen.

3. An immunodiagnostic enhancer as described in claim 1, characterized in that, The pH adjuster may be one or a mixture of the following: disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, sodium citrate, 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), 2-morpholinoethanesulfonic acid (MES), 3-morpholinopropanesulfonic acid (MPOS), 2-[[tris(hydroxymethyl)methyl]amino]ethanesulfonic acid (TES), piperazine-N,N'-di(2-ethanesulfonic acid) (PIPES), tris(hydroxymethyl)aminomethane (Tris), tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl), and tris(hydroxymethyl)aminopropanesulfonic acid (TAPS).

4. An immunodiagnostic enhancer as described in claim 1, characterized in that, The protective protein can be a purified protein, polypeptide, or oligopeptide, preferably one or a mixture of human serum albumin (HSA), bovine serum albumin (BSA), ovalbumin (OVA), gelatin, casein and its derivatives, oligopeptides, polypeptides, fish serum albumin, etc. The preferred protective proteins are HSA, BSA, OVA, casein and its derivatives, and fish serum albumin, and even more preferably HSA, BSA, and fish serum albumin.

5. An immunodiagnostic enhancer as described in claim 1, characterized in that, The surfactant can be one or more of nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants; the nonionic surfactants include polysorbates and alkylphenol polyoxyethylene ethers, such as Tween 20, Tween 40, Tween 60, Tween 80, and Triton, with Tween 20 and Triton being preferred; the anionic surfactants include sulfonate surfactants, with sodium dodecyl sulfonate (SDS) being preferred; the cationic surfactants include quaternary ammonium salt surfactants, including dodecyltrimethylammonium bromide (DTAB), tetradecyltrimethylammonium bromide, and hexadecyltrimethylammonium bromide (CTAB); the amphoteric surfactants include Tetronic 1307 (S9), and more preferably Tween 20, Tween 40, and S9.

6. An immunodiagnostic enhancer as described in claim 1, characterized in that, The inorganic salt can be one or a mixture of common inorganic salts such as sodium chloride, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, sodium citrate, sodium ascorbate, potassium phosphate, sodium phosphate, ammonium sulfate, ammonium chloride, and sodium gluconate. And / or the antibacterial agent can be an inorganic or organic compound with antibacterial activity, such as sodium azide, potassium sorbate, potassium sorbitol, sodium phenylpropionate, isothiazolinone, methylchloroisothiazolinone, methylisothiazolinone, Proclin-300, etc., with preferred antibacterial agents being sodium azide, methylchloroisothiazolinone, methylisothiazolinone, and Proclin-300, and more preferably sodium azide and Proclin-300; Preferably, the immunodiagnostic enhancer includes any one of the following: (1) PB, NaCl, BSA, Tween 20, butanol, sodium azide; (2) MES, NaCl, BSA, Tween 20, butanol, Proclin 300; (3) MES, NaCl, ammonium sulfate, BSA, S9, PEG10000, Proclin 300; (4) PB, NaCl, ammonium sulfate, BSA, Tween 40, PEG20000, Proclin 300; (5) Tris, NaCl, ammonium sulfate, HSA, Tween 40, PEG10000, sodium azide; more preferably, the immunodiagnostic enhancer includes any one of the following: (1) PB, NaCl, BSA, Tween 20, butanol, sodium azide; (2) MES, NaCl, BSA, Tween 20, butanol, Proclin 300; Preferably, the content of the enhancer is 0.01-30%, more preferably 5-15%, and even more preferably 10%; the content of the pH adjuster is 1-1000 mM, more preferably 10-200 mM, and even more preferably 50 mM; the content of the protective protein is 0.01-20%, more preferably 0.5-5%, and even more preferably 1%; the content of the surfactant is 0.01-10%, more preferably 0.05-2%, and even more preferably 0.1%; the content of the inorganic salt is 50 mM-1 M, more preferably 150 mM; and the content of the antibacterial agent is 0.005-5%, more preferably 0.05%. Preferably, the immunodiagnostic enhancer comprises 1-1000mM MES at pH 6.0, 50mM-1M NaCl, 0.01-20% BSA, 0.01-10% Tween 20, 0.01-30% butanol, and 0.005-5% Proclin 300.

7. A method for preparing the immunodiagnostic enhancer, comprising the step of mixing an enhancer, a pH adjuster, a protective protein, a surfactant, an inorganic salt, and an antibacterial agent.

8. A method for improving the efficiency of an immunodiagnostic reaction, comprising the step of adding the immunodiagnostic enhancer according to any one of claims 1-6 to an immunodiagnostic reaction system, wherein, The immunodiagnostic reaction is selected from enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (chemiluminescence immunoassay), or immunochromatographic reaction. Preferably, the immunodiagnostic enhancer is added at a ratio of 10% in the immunodiagnostic reaction system; Preferably, in the enzyme-linked immunosorbent assay (ELISA) reaction, the immunodiagnostic enhancer is added to a 96-well plate, followed by the addition of sample, enzyme conjugate, and substrate for incubation and color development; in the chemiluminescence reaction, the immunodiagnostic enhancer replaces the solvent of the magnetic bead components; and in the immunochromatographic reaction, the immunodiagnostic enhancer is added to the sample buffer to form a mixed dilution. Preferably, after using the immunodiagnostic enhancer, the detection signal intensity is increased by at least twice compared to the control group without the enhancer.

9. Use of the immunodiagnostic enhancer according to any one of claims 1-6 in the preparation of reagents that improve the efficiency of immunodiagnostic reactions, wherein, The immunodiagnostic reaction is selected from enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (chemiluminescence immunoassay), or immunochromatographic reaction. Preferably, the improvement in the efficiency of the immunodiagnostic reaction is manifested in improving the antigen-antibody binding efficiency and / or increasing the intensity of the detection signal, including OD value, relative luminescence value or fluorescence intensity.

10. An immunodiagnostic kit, comprising the immunodiagnostic enhancer as described in any one of claims 1-6, and one or more of enzyme-linked immunosorbent assay (ELISA) reagents, chemiluminescent reagents, or immunochromatographic reagents; Preferably, the kit further includes a sample diluent, magnetic bead components, an enzyme conjugate, a substrate, or a sample buffer, wherein, The immunodiagnostic enhancer is used in combination with the sample diluent, magnetic bead components, or sample buffer.

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