Modified luminous particle as well as preparation method and application thereof

Abstract

The invention relates to a modified luminous particle as well as a preparation method and application thereof. The modified light-emitting particles comprise light-emitting particles and zwitterionic polymers adsorbed on the light-emitting particles, and the modified light-emitting particles can react with active oxygen to generate detectable chemiluminescence signals. According to the scheme provided by the invention, the repulsive force between particles and the non-specific adsorption resistance can be improved, so that the specificity of the light-excited chemiluminescence detection reagent is improved, and the accuracy of a detection result is ensured.

Description

Technical Field

[0001] The present application relates to the field of immunoassay technology, and in particular to a modified luminescent particle and a preparation method and application thereof. Background Art

[0002] Immunoassays are a widely used clinical testing method. They utilize the specific reaction between antigens and antibodies to identify the analyte, and then utilize the signal generated by the recognized analyte with a reagent for quantitative analysis. Depending on whether the analyte is separated from the reaction system during the assay, immunoassays can be categorized into heterogeneous and homogeneous phases. Light-initiated chemiluminescence immunoassays are a typical homogeneous immunoassay technique.

[0003] The photochemiluminescence detection system consists of photosensitive particles and luminescent particles. The photosensitive particles are doped with photosensitizers that generate reactive oxygen species when excited by light, while the luminescent particles are doped with dyes that react with reactive oxygen species to produce fluorescence. When the molecule to be detected is present, it forms an immune complex with the luminescent particles coupled with the capture antigen / antibody and the photosensitive particles coupled with the detection antigen / antibody through a double antibody sandwich effect. The luminescent particles then receive the reactive oxygen species produced by the photosensitive particles and stimulate a light signal. When the molecule to be detected is absent, the immune complex cannot form between the two particles. The distance between the luminescent and photosensitive particles exceeds the propagation range of the reactive oxygen species, and no light signal is generated during detection. The photochemiluminescence detection method has the characteristics of high sensitivity, no washing required, good reproducibility, and simple and easy operation. At the same time, because the detection process does not involve a separation and washing step, the detection results are easily interfered with by various factors present in the sample. For example, impurities in the sample that bind to the luminescent particles will affect the detection specificity and, in turn, the accuracy of the detection results. Summary of the Invention

[0004] In order to solve or partially solve the problems existing in the related art, the present application provides a modified luminescent particle and its preparation method and application, which can improve the repulsive force between the luminescent particles and the ability to resist nonspecific adsorption, thereby improving the specificity of the photochemiluminescence detection reagent and ensuring the accuracy of the detection results.

[0005] In a first aspect, the present application provides a modified luminescent particle, comprising a luminescent particle and a zwitterionic polymer adsorbed on the luminescent particle, wherein the modified luminescent particle can react with active oxygen to generate a detectable chemiluminescent signal.

[0006] In some embodiments of the present application, the zwitterionic polymer is formed by polymerizing a single zwitterionic polymer monomer.

[0007] In some embodiments of the present application, the average molecular weight of the zwitterionic polymer is 1000 to 60000 g / mol.

[0008] In some embodiments of the present application, the modified luminescent microparticle further comprises a biodetection molecule coupled to the surface of the luminescent microparticle, and the biodetection molecule can specifically bind to the molecule to be detected.

[0009] In some embodiments of the present application, when the biological detection molecule is an antigen, the molecular weight of the zwitterionic polymer is preferably 1000 to 20000 g / mol.

[0010] In some embodiments of the present application, when the biodetection molecule is an antibody, the molecular weight of the zwitterionic polymer is preferably 20,000 to 60,000 g / mol.

[0011] In some embodiments of the present application, the anionic group of the zwitterionic polymer is selected from carboxylic acid anions, sulfonic acid anions, or phosphoric acid anions.

[0012] In some embodiments of the present application, the cationic group of the zwitterionic polymer is selected from quaternary ammonium salt ions, phosphonic acid groups, pyridine groups or imidazole groups.

[0013] In some preferred embodiments of the present application, the zwitterionic polymer is selected from one or more of polymethacryloyloxyethyl phosphorylcholine, polysulfobetaine methacrylate, and polycarboxybetaine methacrylate; preferably selected from polysulfobetaine methacrylate or polycarboxybetaine methacrylate.

[0014] In some embodiments of the present application, the average increase in particle size of the luminescent particles after modification is no more than 10%;

[0015] Preferably, the particle size of the luminescent particles before and after modification is 150 to 300 nm.

[0016] A second aspect of the present application provides a method for preparing modified luminescent particles, comprising the following steps: adding a zwitterionic polymer to a solution containing luminescent particles, allowing the zwitterionic polymer to adsorb on the luminescent particles, to obtain modified luminescent particles.

[0017] The third aspect of the present application provides a photochemiluminescence detection reagent, comprising the modified luminescent particles as described in the first aspect of the present application or the modified luminescent particles prepared by the preparation method described in the second aspect of the present application.

[0018] In some embodiments of the present application, the concentration of the zwitterionic polymer in the detection reagent is 0.01 μM to 0.1 μM.

[0019] The fourth aspect of the present application provides a photochemiluminescence detection kit, comprising the photochemiluminescence detection reagent described in the third aspect of the present application.

[0020] The fifth aspect of the present application provides a modified luminescent particle as described in the first aspect of the present application, or the modified luminescent particle prepared by the preparation method described in the second aspect of the present application, or the photochemiluminescence detection reagent described in the third aspect of the present application, or the kit described in the fourth aspect of the present application in photochemiluminescence detection.

[0021] The technical solution provided by the present application may include the following beneficial effects: by adding a zwitterionic polymer to the luminescent reagent component of the photochemiluminescent antigen (or antibody) detection reagent, the zwitterionic polymer is adsorbed onto the surface of the luminescent particles of the luminescent reagent, thereby forming modified luminescent particles and a modified photochemiluminescent detection reagent. In the modified photochemiluminescent detection reagent, due to the hydration effect of the zwitterionic polymer, a free energy barrier is formed, and at the same time, the zwitterionic polymer has the characteristics of charge balance and dipole minimization, which increases the repulsive force between the modified luminescent particles, avoids nonspecific adsorption of the particles, thereby improving the specificity of the detection reagent and ensuring the accuracy of the detection results.

[0022] Furthermore, the zwitterionic polymers, modified luminescent particles, photochemiluminescence detection reagents, etc. in this scheme are easy to prepare, highly operational, and have a significant effect on improving detection specificity, which is conducive to their application in photochemiluminescence detection systems. The detection reagents have a wide range of applications and high accuracy of detection results, and are worth promoting to other chemiluminescence detection projects. DETAILED DESCRIPTION

[0023] To facilitate understanding of the present invention, the present invention will be described in detail below. However, before describing the present invention in detail, it should be understood that the present invention is not limited to the specific embodiments described. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting.

[0024] Where a range of values is provided, it is understood that each intervening value between the upper and lower limits of the range and any other specified or intervening values in the specified range is encompassed within the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the present invention, subject to any express exclusions in the specified range. Where a specified range includes one or both limits, ranges excluding either or both of those included limits are also encompassed within the present invention.

[0025] Unless otherwise defined, all terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although any methods and materials or equivalents to those described herein may also be used in the practice or testing of the present invention, preferred methods and materials are now described.

[0026] I. Terminology

[0027] The term "test molecule" as used herein refers to a substance in a sample to be tested. The test molecule may be, for example, a polypeptide, protein, polysaccharide, glycoprotein, nucleic acid, or small molecule compound.

[0028] As used herein, the term "sample" refers to a mixture containing or suspected of containing the molecule to be tested. Samples may include body fluids such as blood, plasma, serum, urine, semen, saliva, cell culture, and tissue extracts. Samples may be diluted with a diluent as needed before use. For example, to avoid the "hook effect," the sample may be diluted with a diluent before testing on the instrument.

[0029] As used herein, the term "particle" refers to a small, localized object that can be attributed to physical properties such as volume, mass, or size. Particles can be symmetrical, spherical, or irregular, asymmetrical in shape; they can be of any size. Particles suitable for use herein can be spherical, for example, particles with diameters in the nanometer or micrometer range. Particles can be solids (e.g., polymers, metals, glasses, organic or inorganic materials such as minerals, salts, and diatoms), oil droplets (e.g., hydrocarbons, fluorocarbons, siliceous fluids), or vesicles (e.g., synthetic such as phospholipids, or natural such as cells and organelles). Particles can be organic or inorganic, such as latex particles or other particles containing organic or inorganic polymers, lipid bilayers such as liposomes, phospholipid vesicles, oil droplets, silica particles, metal sols, cells, and microcrystalline dyes. Particles can be expandable or non-expandable, porous or non-porous, have any density, but preferably have a density close to that of water, preferably float in water, and be composed of a transparent, partially transparent, or opaque material.

[0030] The term "antibody" as used herein is used in the broadest sense and includes antibodies of any isotype, antibody fragments that retain specific binding to an antigen, including but not limited to Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies, bispecific antibodies, and fusion proteins comprising an antigen-binding portion of an antibody and a non-antibody protein. Where necessary, the antibody may be further conjugated to other moieties, such as biotin or avidin.

[0031] As used herein, the term "antigen" refers to a substance that can stimulate a substrate to produce an immune response and bind to the immune response products, antibodies, and sensitized lymphocytes, both in vivo and in vitro, to produce an immune effect. Antigens include, but are not limited to, bacteria, viruses, bacterial exotoxins, and most proteins. Where desired, antigens may be further conjugated to other moieties, such as biotin or avidin.

[0032] As used herein, the term "binding" refers to the direct association between two molecules due to interactions such as covalent, electrostatic, hydrophobic, ionic and / or hydrogen bonds, including but not limited to interactions such as salt bridges and water bridges.

[0033] The term "specific binding" as used herein refers to the mutual recognition and selective binding reaction between two substances, which refers to the conformational correspondence between the responding reactants from a three-dimensional structural perspective.

[0034] The term "active oxygen species" as used herein refers to a general term for substances in the body or in the natural environment that are composed of oxygen, contain oxygen, and are active in nature, mainly an excited state of oxygen molecules, including the one-electron reduction product of oxygen, superoxide anion (O2·-), the two-electron reduction product of oxygen, hydrogen peroxide (H2O2), the three-electron reduction product of oxygen, hydroxyl radical (·OH), as well as nitric oxide and reactive oxygen species ( 1 O2) etc.

[0035] II. Specific implementation plan

[0036] The present application will be described in more detail below.

[0037] The main evaluation indicators of immunoassay systems are sensitivity and specificity. Sensitivity refers to the ability of the assay system to detect extremely low levels of the analyte, reflecting the system's ability to detect the analyte. Specificity refers to the ability of the assay system to detect a specific analyte, reflecting the system's ability to correctly identify the analyte.

[0038] The specificity of immunoassay systems is susceptible to interference from various factors present in the sample, such as sample quality, reagent quality, improper operation, cross-reactions, and instrumentation. Poor sample quality, such as impurities or excessive protein content in blood samples, can interfere with the accuracy of immunoassay results. Poor reagent quality, such as impure or expired antibodies or antigens, can lead to false-positive or false-negative results. Improper operation, such as inaccurate sample volume or excessively long or short incubation times, can also affect specificity. Cross-reactions between certain antigens and antibodies can lead to false-positive results. Inaccurate or unstable instrumentation can also reduce specificity.

[0039] Different immunoassay platforms have different factors that affect the specificity of the corresponding detection system. Photochemiluminescence detection reagents include luminescent reagents, photosensitive reagents, etc. For the photochemiluminescence detection platform, the applicant found that the specificity of the luminescent reagent will also directly affect the detection results fed back by the detection system. By improving the ability of the luminescent reagent in the detection reagent to resist nonspecific adsorption, the probability of impurities in the sample binding to the luminescent particles can be reduced, which is a feasible method to improve the specificity of the detection system.

[0040] The modified luminescent particles involved in the first aspect of the present invention comprise luminescent particles and a zwitterionic polymer adsorbed onto the luminescent particles. The modified luminescent particles are capable of reacting with reactive oxygen species to produce a detectable chemiluminescent signal. The luminescent particles are doped with a chemiluminescent agent and a fluorescent agent. The chemiluminescent agent converts the energy of the reactive oxygen species into 360nm emitted light, which excites the fluorescent agent to produce fluorescence. The chemiluminescent agent can be selected from dimethylthiophene; the fluorescent agent can be selected from one or more lanthanide metal complexes such as europium, terbium, and samarium.

[0041] The modified luminescent particles of the present application can be used to prepare photo-induced chemiluminescence detection reagents after being mixed with chemiluminescent agents and fluorescent agents.

[0042] The zwitterionic polymers described in the examples of this application are a new generation of anti-adsorption materials. They are a class of polymers with a pair of oppositely charged atoms within their repeating units. Modified luminescent microparticles adsorbed with zwitterionic polymers can significantly enhance interparticle repulsion and prevent nonspecific adsorption, thereby improving the specificity of immunoassays and enhancing the accuracy of test results.

[0043] Zwitterionic polymers have the following characteristics: (1) balanced charge, with equal amounts of positive and negative charged groups; (2) minimized dipole, with even distribution of positive and negative charges at the molecular level; and (3) strong hydration effect. Compared to the hydration effect of traditional nonionic hydrophilic materials through hydrogen bonding, zwitterionic polymers can undergo stronger hydration through ion solvation. This zwitterionic polymer material with strong hydration effect, charge balance, and minimized dipole properties is adsorbed on luminescent microparticles to modify the luminescent microparticles, making the modified luminescent microparticles have excellent anti-bioadhesion properties, including anti-nonspecific protein adsorption and anti-cell adhesion. Therefore, the modified luminescent microparticles can maintain high stability in highly complex media (such as undiluted serum) and will not electrostatically adsorb proteins due to local charge generation. At the same time, due to the strong hydration effect, a free energy barrier is formed, which can increase the repulsive force between the microparticles, thereby improving the nonspecific adsorption ability of the luminescent microparticles as a whole and improving the specificity of the photoinduced chemiluminescent antigen / antibody detection reagent.

[0044] In some embodiments, the zwitterionic polymer comprises equal amounts of anionic groups and cationic groups such that the sum of the charges of the zwitterionic polymer is zero, wherein the anionic groups can be selected from carboxylic acid anions, sulfonic acid anions, or phosphate anions, and the cationic groups can be selected from quaternary ammonium ions, phosphonic acid groups, pyridinium groups, or imidazole groups.

[0045] In some preferred embodiments, the zwitterionic polymer is selected from one or more of polymethacryloyloxyethylphosphorylcholine (polyMPC), polysulfobetaine methacrylate (polySBMA), and polycarboxybetaine methacrylate (polyCBMA); more preferably, polysulfobetaine methacrylate or polycarboxybetaine methacrylate.

[0046] Among them, polysulfobetaine methacrylate is prepared by polymerizing its monomer sulfobetaine methacrylate SBMA, and polycarboxybetaine methacrylate is prepared by polymerizing its monomer carboxybetaine methacrylate CBMA.

[0047] In some embodiments, the zwitterionic polymer is polymerized from a single zwitterionic polymer monomer. The zwitterionic polymer can be polymerized using a free radical polymerization (APS) method. Furthermore, the initiator used in the polymerization of the zwitterionic polymer is selected from ammonium persulfate.

[0048] During the synthesis of zwitterionic polymers, by adjusting the ratio of monomer to initiator, polymers with different degrees of polymerization, i.e., polymers with different average molecular weights, can be obtained. In practical applications, polymers with different average molecular weights can be selected as needed.

[0049] In some embodiments, the average molecular weight of the zwitterionic polymer is 1,000 to 60,000 g / mol.

[0050] In some embodiments, the modified luminescent microparticles further include biodetection molecules coupled to the surface of the luminescent microparticles, wherein the biodetection molecules can specifically bind to the molecules to be detected. For example, the biodetection molecules can be antibody molecules or antigen molecules.

[0051] Specifically, during detection, the modified luminescent microparticles bind to the molecule being tested via the biodetection molecules bound to their surfaces. When the molecule being tested is an antibody, the biomolecule bound to the surface of the modified luminescent microparticles in the detection reagent is an antigen, which binds to the molecule being tested through specific antigen-antibody binding. When the molecule being tested is an antigen, the biomolecule bound to the surface of the modified luminescent microparticles in the detection reagent is an antibody, which binds to the molecule being tested through specific antigen-antibody binding.

[0052] In some specific embodiments, when the detection item is antibody detection, that is, the molecule to be detected is an antibody, the modified luminescent particles are coupled (coated) with an antigen, and the molecular weight of the zwitterionic polymer is preferably 1000-20000 g / mol; more preferably, it is polysulfobetaine methacrylate with a molecular weight of 1000-20000 g / mol; further preferably, it is polysulfobetaine methacrylate with a molecular weight of 1000-15000 g / mol.

[0053] When the biological detection molecule is an antibody, that is, the molecule to be detected is an antigen, the molecular weight of the zwitterionic polymer is preferably 20,000 to 60,000 g / mol.

[0054] In some specific embodiments, when the detection item is antigen detection, that is, the molecule to be detected is an antigen, the modified luminescent particles are coupled (coated) with antibodies, and the molecular weight of the zwitterionic polymer is preferably 20,000 to 60,000 g / mol; more preferably, it is polycarboxybetaine methacrylate with a molecular weight of 20,000 to 60,000 g / mol; further preferably, it is polycarboxybetaine methacrylate with a molecular weight of 25,000 to 60,000 g / mol.

[0055] In some embodiments, the average increase in particle size of the luminescent particles after modification is no greater than 10%.

[0056] Preferably, the particle size of the luminescent particles before and after modification is 150 to 300 nm. After the luminescent particles are adsorbed with the zwitterionic polymer, the particle size does not change significantly.

[0057] The change in particle size of the luminescent microparticles before and after modification can be determined by performing an elution test on the modified luminescent microparticles and calculating the elution rate of the modified luminescent microparticles to obtain the change in particle size before and after modification, thereby demonstrating the effect of the zwitterionic polymer on the nonspecific protein adsorption ability of the microparticles. The elution test includes: first, taking luminescent microparticles from a solution containing luminescent microparticles to determine their initial particle size; after adding the zwitterionic compound to the solution containing the luminescent microparticles, the solution is allowed to stand at low temperature for a period of time, and then sampling to determine the particle size of the modified luminescent microparticles before elution; then, the modified luminescent microparticles are centrifuged and resuspended in pure water for washing to obtain the eluted modified luminescent microparticles, and then sampling to determine the particle size of the modified luminescent microparticles after elution and calculate the elution rate.

[0058]

[0059] The embodiment of the present application also relates to a method for preparing modified luminescent particles, comprising the following steps: adding a zwitterionic polymer to a solution containing luminescent particles, so that the zwitterionic polymer is adsorbed on the luminescent particles to obtain modified luminescent particles.

[0060] In some embodiments, the solution containing luminescent particles may refer to a luminescent reagent in a detection reagent, which includes luminescent particles and a buffer. The luminescent particles are suspended in the buffer, and the luminescent particles can react with reactive oxygen species to produce a chemiluminescent signal. The buffer may be selected from, for example, HEPES buffer, Tris-HCl buffer, or PBS buffer.

[0061] The embodiments of the present application also relate to a photo-induced chemiluminescence detection reagent, comprising the modified luminescent particles described above or the modified luminescent particles prepared by the preparation method of the modified luminescent particles described above.

[0062] In some embodiments, the concentration of the zwitterionic polymer in the detection reagent is 0.01 μM to 0.1 μM, such as 0.03 μM, 0.05 μM, 0.10 μM or any other concentration value within the above range.

[0063] The preparation method of the photoluminescence detection reagent comprises the following steps:

[0064] A zwitterionic polymer with a concentration of 0.01 μM to 0.1 μM is added to a luminescent reagent containing luminescent particles, and the zwitterionic polymer is adsorbed onto the surface of the luminescent particles to modify the luminescent particles to obtain modified luminescent particles, thereby preparing a modified photochemiluminescence detection reagent.

[0065] The method for detecting a molecule to be tested of the present application comprises the following steps:

[0066] (1) mixing the sample with the modified photochemiluminescence detection reagent and incubating the mixture;

[0067] (2) Detecting the optical signal of the mixture obtained in step (1).

[0068] The present application also relates to a photochemiluminescence detection kit, comprising the above-mentioned photochemiluminescence detection reagent.

[0069] The present application also relates to the use of the modified luminescent particles, photochemical detection reagents or photochemical detection kits in photochemiluminescence detection.

[0070] The molecular detection methods used in this application are for non-diagnostic or therapeutic purposes.

[0071] III. Specific Examples

[0072] To make the present invention easier to understand, the present invention will be further described in detail below with reference to the following examples. These examples are merely illustrative and do not limit the scope of application of the present invention. Unless otherwise specified, the raw materials or components used in the present invention can be obtained through commercial channels or conventional methods.

[0073] Example 1

[0074] 1. Experimental materials:

[0075] SBMA monomer, CBMA monomer, ammonium persulfate, anhydrous ethanol, deionized water.

[0076] 2. Experimental methods:

[0077] SBMA monomer and initiator ammonium persulfate were mixed and dissolved in deionized water, and the molar ratio of monomer raw material to initiator was adjusted. The mixture was reacted at 70°C for 6 hours, cooled at 4°C for 3 hours, and then slowly added to anhydrous ethanol. The mixture was then washed with deionized water several times to obtain polySBMA with different polymerization degrees.

[0078] The CBMA monomer and the initiator ammonium persulfate were mixed and dissolved in deionized water, and the molar ratio of the monomer raw material to the initiator was adjusted. The mixture was reacted at 70°C for 6 hours, and then cooled at 4°C for 3 hours. Then, the mixture was slowly added to anhydrous ethanol and washed with deionized water several times to obtain polyCBMA with different polymerization degrees.

[0079] 3. Experimental data:

[0080] Table 1 SBMA and CBMA monomer polymerization results

[0081]

[0082] 4. Experimental conclusion:

[0083] Using the free radical polymerization method (APS method), polySBMA and polyCBMA with different polymerization degrees were synthesized from SBMA and CBMA monomers, respectively, with an average molecular weight ranging from 1000 to 60000 g / mol.

[0084] Example 2

[0085] 1. Experimental materials:

[0086] Anti-HCV detection kit (lot number: L2211), HBeAg detection kit (lot number: L2203), polySBMA, polyCBMA.

[0087] 2. Experimental methods:

[0088] Select the analytes as antigens and antibodies Use one detection kit for each test, take an appropriate amount of luminescent reagent (R1 reagent), divide the R1 reagent into 4mL / group x18 groups, dilute each group to 10μg / mL, and use a particle size analyzer to measure the initial particle size. Then add the zwitterionic polymers of different concentrations and different polymerization degrees prepared in Example 1 to each group to prepare a modified luminescent reagent. After standing at 4°C for 24h, take a sample to measure the particle size of the modified luminescent particles before elution. Then centrifuge the modified luminescent reagent at 15000rpm for 20min, resuspend and wash with pure water, repeat 3 times, take a sample to measure the particle size of the modified luminescent particles after elution, and calculate the elution rate.

[0089]

[0090] 3. Experimental data:

[0091] Table 2 Effect of polySBMA on the particle size of Anti-HCV luminescent particles

[0092]

[0093] Table 3 Effect of polyCBMA on the particle size of HBsAg luminescent particles

[0094]

[0095]

[0096] 4. Experimental conclusion:

[0097] When zwitterionic polymers with different polymerization degrees were added to the R1 reagent, the particle size of the luminescent particles increased to varying degrees after standing for a period of time, and increasing the concentration did not affect the increase in the particle size after 24 hours (particle size before elution).

[0098] After adding polySBMA or polyCBMA and allowing the particles to stand for 24 hours, the average particle size increase was 5% and 9%, respectively. After 24 hours of standing, the particles were centrifuged and washed, and the particle size was measured again, showing close to the initial size (P-Values of 0.14805 and 0.323836, respectively), indicating that the adsorbed zwitterionic polymer was eluted. This indicates that after adding the zwitterionic polymer, the luminescent particles adsorbed the polymer, and the particle size stabilized after 24 hours of standing.

[0099] Example 3

[0100] 1. Experimental materials:

[0101] Anti-HCV detection kit (lot number: L2211), HBeAg detection kit (lot number: L2203), polySBMA, polyCBMA, serum (Anti-HCV negative, HBeAg negative), purified water.

[0102] 2. Experimental methods:

[0103] A modified luminescent reagent (0.05 μM) was prepared from each experimental group prepared in Example 1, using the method of Example 2. A control group consisted of a luminescent reagent prepared by adding an equal volume of purified water and incubating for 24 hours. All groups were mixed with serum at a 1:1 ratio and incubated at 4°C for 24 hours. The particle size was then measured and compared with that of the control group.

[0104] 3. Experimental data:

[0105] Table 4 Effect of polySBMA on nonspecific adsorption of Anti-HCV anti-protein

[0106]

[0107] Table 5 Effect of polyCBMA on nonspecific adsorption of HBeAg anti-protein

[0108]

[0109]

[0110] 4. Experimental conclusion:

[0111] After the luminescent particles were mixed with serum and adsorbed for 24 hours, the change rate of the modified group was lower than that of the control group, and the protein adsorption level was lower than that of the control group.

[0112] For anti-HCV (antibody testing) testing, polySBMA with a molecular weight between 1,000 and 15,000 exhibited superior anti-adsorption properties; for HBeAg testing, polyCBMA with a molecular weight between 25,000 and 60,000 exhibited superior anti-adsorption properties. Considering the differences in the antibodies and antigens coated on the luminescent microparticles for the two testing methods, these results suggest that for testing with antigens coated on the luminescent microparticles, zwitterionic polymers with a molecular weight between 1,000 and 20,000 may exhibit superior anti-adsorption properties; for testing with antibodies coated on the luminescent microparticles, zwitterionic polymers with a molecular weight between 20,000 and 60,000 may exhibit superior anti-adsorption properties.

[0113] Example 4

[0114] 1. Experimental materials:

[0115] Anti-HCV detection kit (lot number: L2211), HBeAg detection kit (lot number: L2203), polySBMA, polyCBMA, and clinical samples with interference (anti-HCV negative, HBeAg negative)

[0116] 2. Experimental methods:

[0117] The group with better anti-adsorption effect in Example 2 was selected to replace the luminescent reagent in the kit to assemble an experimental reagent group. The original reagent in the kit was used as a comparison to detect clinical samples. The detection concentration values (S / CO) were compared. If S / CO ≥ 1, the detection sample was judged to be positive, and if S / CO < 1, the detection sample was judged to be negative.

[0118] 3. Experimental data:

[0119] Table 6 Effect of polySBMA on Anti-HCV detection specificity

[0120]

[0121] Table 7 Effect of polyCBMA on HBeAg detection specificity

[0122]

[0123]

[0124] 4. Experimental conclusion:

[0125] Compared with the control luminescent reagent, the modified luminescent reagent doped with zwitterionic polymer can significantly reduce the measurement values of clinical samples (Anti-HCV and HBeAg) with interference, thereby improving Improve the specificity of antibody or antigen detection reagents and reduce the false positive rate.

[0126] It should be noted that the embodiments described above are only used to explain the present application and do not constitute any limitation to the present application. The present application has been described with reference to typical embodiments, but it should be understood that the words used therein are descriptive and explanatory words, rather than restrictive words. The present application may be modified as specified within the scope of the claims of the present application, and the present application may be revised without departing from the scope and spirit of the present application. Although the present application described therein relates to specific methods, materials and embodiments, it does not mean that the present application is limited to the specific examples disclosed therein. On the contrary, the present application can be extended to all other methods and applications with the same function.

Claims

1. A modified luminescent particle, characterized in that: The modified luminescent particles include luminescent particles and zwitterionic polymers adsorbed on the luminescent particles. The modified luminescent particles can react with active oxygen to generate detectable chemiluminescent signals.

2. The modified luminescent particle according to claim 1, characterized in that The zwitterionic polymer is formed by polymerization of a single zwitterionic polymer monomer; Preferably, the average molecular weight of the zwitterionic polymer is 1000 to 60000 g / mol.

3. The modified luminescent particle according to claim 1 or 2, characterized in that: The modified luminescent microparticles further include biological detection molecules coupled to the surface of the luminescent microparticles, wherein the biological detection molecules are capable of specifically binding to the molecules to be detected; When the biodetection molecule is an antigen, the molecular weight of the zwitterionic polymer is preferably 1000 to 20000 g / mol; When the biodetection molecule is an antibody, the molecular weight of the zwitterionic polymer is preferably 20,000 to 60,000 g / mol.

4. The modified luminescent particle according to claim 1, characterized in that The anionic group of the zwitterionic polymer is selected from carboxylic acid anions, sulfonic acid anions or phosphoric acid anions, and the cationic group is selected from quaternary ammonium salt ions, phosphonic acid groups, pyridine groups or imidazole groups; Preferably, the zwitterionic polymer is selected from polysulfobetaine methacrylate and polycarboxybetaine methacrylate.

5. The modified luminescent particle according to claim 1, characterized in that The average increase in the particle size of the luminescent particles after modification is no more than 10%; Preferably, the particle size of the luminescent particles before and after modification is 150 to 300 nm.

6. A method for preparing the modified luminescent particles according to any one of claims 1 to 5, characterized in that: The steps include: A zwitterionic polymer is added to a solution containing luminescent fine particles, and the zwitterionic polymer is adsorbed on the luminescent fine particles to obtain modified luminescent fine particles.

7. A photochemiluminescence detection reagent, characterized in that: The modified luminescent particles include the modified luminescent particles according to any one of claims 1 to 5 or the modified luminescent particles prepared by the preparation method according to claim 6.

8. The photochemiluminescence detection reagent according to claim 7, characterized in that The concentration of the zwitterionic polymer in the detection reagent is 0.01 μM to 0.1 μM.

9. A photochemiluminescence detection kit, characterized in that: Comprising the photochemiluminescence detection reagent as described in claim 7 or 8.

10. Use of the modified luminescent microparticles according to any one of claims 1 to 5, or the modified luminescent microparticles prepared by the preparation method according to claim 6, or the detection reagent according to claim 7 or 8, or the kit according to claim 9 in photochemiluminescence detection.

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