Magnetic particle, magnetic particle coating substance and preparation method and application of magnetic particle coating substance

Abstract

The invention relates to the technical field of biomedicine, in particular to a magnetic particle, a magnetic particle coating substance and a preparation method and application of the magnetic particle coating substance. The magnetic particle coating substance comprises a magnetic particle and a hepatitis B virus protein antibody coated on the surface of the magnetic particle, and the surface of the magnetic particle is modified with L-valine-D8; and / or, the protein antibody in the hepatitis B virus is modified by reacting with N-hydroxysuccinimide ester and pyridyl disulfide in advance. When the magnetic particle or the magnetic particle coating substance is used for carrying out chemiluminescence immunodetection on the protein in the hepatitis B virus surface antigen, the background can be reduced, the signal intensity can be improved and the like, and the magnetic particle or the magnetic particle coating substance has good detection performance.

Description

TECHNICAL FIELD

[0001] The present application relates to the biomedical technology field, and in particular to a magnetic microparticle, a magnetic microparticle coating and a preparation method and application thereof. BACKGROUND

[0002] Immune detection as an important biological analysis method, plays a key role in medical diagnosis, drug research and development, food safety and other fields. And magnetic bead technology, especially immune magnetic bead (IMB) technology, with its unique performance and wide application prospect, plays an increasingly important role in immune detection.

[0003] Immune magnetic beads are small spherical particles that combine immunology and magnetism principles, usually composed of a magnetic core, a polymer coating and a functional base layer. The magnetic core endows the magnetic beads with superparamagnetism, allowing them to be quickly separated under the action of an external magnetic field; the polymer coating is used to stabilize the surface of the magnetic beads and prevent aggregation; the functional base layer is chemically or biologically modified to bind specific antibodies or antigens, enabling the capture and separation of target molecules.

[0004] Middle hepatitis B surface antigen (MHBs) is an important component of hepatitis B virus (HBV) surface antigen. MHBs is an important serological marker of HBV infection, and its detection has important clinical value. However, in the prior art, the conventional magnetic bead coating coupling process route cannot meet the detection requirements of MHBs. SUMMARY

[0005] The purpose of the present application is to provide a magnetic microparticle, a magnetic microparticle coating and a preparation method and application thereof, which has good detection performance when used for chemiluminescence immunoassay of middle hepatitis B surface antigen.

[0006] To this end, in a first aspect, the present application provides a magnetic microparticle, the surface of which is modified with L-valine-D8.

[0007] In a second aspect of the present application, a preparation method of the magnetic microparticle is provided, which comprises:

[0008] A magnetic microparticle is provided, and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysulfosuccinimide (Sulfo-NHS) are used to perform a first activation treatment on the magnetic microparticle; then L-valine-D8 is modified on the surface of the magnetic microparticle.

[0009] In some embodiments, in the first activation treatment, the mass ratio of EDC to Sulfo-NHS is 1.5-2:1.

[0010] In some embodiments, the reaction conditions for the first activation treatment and / or the modification of L-valine-D8 include: rotating the reaction at room temperature for 30 to 120 minutes.

[0011] The third aspect of the present invention provides a magnetic particle coating, comprising magnetic particles and hepatitis B virus middle protein (MHBs) antibodies coated on the surface of the magnetic particles; wherein,

[0012] The surface of the magnetic particles is modified with L-valine-D8; and / or,

[0013] The MHBs antibody was previously modified by reacting with the following groups: N-hydroxysuccinimide ester, pyridyl disulfide.

[0014] In some embodiments, the method for modifying the surface of the magnetic microparticles with L-valine-D8 comprises: providing magnetic microparticles, performing a first activation treatment on the magnetic microparticles with EDC and Sulfo-NHS; and then modifying the surface of the magnetic microparticles with L-valine-D8.

[0015] In some embodiments, the MHBs antibody is pre-modified by reacting with sulfosuccinimidyl 6-(3'-(2-pyridyldithio)propionamido)hexanoate (Sulfo-LC-SPDP).

[0016] A fourth aspect of the present invention provides a method for preparing the magnetic particle coating, comprising:

[0017] The magnetic particles are subjected to a second activation treatment using EDC and Sulfo-NHS to obtain activated magnetic particles; the MHBs antibody is mixed with the activated magnetic particles to perform a coating reaction to prepare the magnetic particle coating.

[0018] In some embodiments, the surface of the magnetic microparticles is modified with L-valine-D8, and the L-valine-D8 is modified by the following steps:

[0019] Magnetic particles are provided, and EDC and Sulfo-NHS are used to perform a first activation treatment on the magnetic particles; and then L-valine-D8 is modified on the surface of the magnetic particles.

[0020] In some embodiments, in the first activation treatment, the mass ratio of EDC to Sulfo-NHS is 1.5 to 2:1.

[0021] In some embodiments, the reaction conditions for the first activation treatment and / or the modification of L-valine-D8 include: rotating the reaction at room temperature for 30 to 120 minutes.

[0022] In some embodiments, the MHBs antibody is pre-modified by reacting with Sulfo-LC-SPDP.

[0023] In some embodiments, after the coating reaction, the following steps are further included: curing reaction;

[0024] The curing reaction includes adding EDC and mixing at room temperature for 30 to 60 minutes. For example, an EDC solution dissolved in pre-cooled (eg, refrigerated at 4° C.) ultrapure water can be added, and the concentration can be about 1 to 5 mg / mL.

[0025] In a fifth aspect, the present invention provides a detection kit for hepatitis B virus protein, which comprises the magnetic particle coating described in the third aspect of the present invention, or the magnetic particle coating prepared according to the preparation method described in the fourth aspect of the present invention.

[0026] The sixth aspect of the present invention provides use of the magnetic particles, magnetic particle coatings or the detection kit for preparing immunoassay products.

[0027] In some embodiments, the immunoassay product is used to detect proteins in hepatitis B virus.

[0028] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0029] The present invention provides hydrophobicized magnetic microparticles with L-valine-D8 modified on the surface, which are used for chemiluminescent immunoassays, and can significantly improve the intensity of the chemiluminescent signal, which is beneficial to improving the sensitivity of the detection. Furthermore, the present invention provides modified hepatitis B protein antibodies for the detection of hepatitis B protein antibodies, which are coated on the surface of the aforementioned hydrophobicized magnetic microparticles, which can simultaneously reduce the background and increase the chemiluminescent signal intensity, thus better meeting the needs of clinical detection. Moreover, after the magnetic microparticles coated with hepatitis B protein antibodies are solidified, their storage stability can be significantly improved, which has excellent practical application prospects. DETAILED DESCRIPTION

[0030] The exemplary embodiments of the present disclosure will be described in more detail below. It should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments set forth herein. Instead, these embodiments are provided to enable a more thorough understanding of the present disclosure and to fully convey the scope of the present disclosure to those skilled in the art.

[0031] Hepatitis B virus surface antigen (HBs) is a key component of the hepatitis B virus (HBV) surface antigen. MHBs is an important serological marker for HBV infection, and its detection has significant clinical value. However, conventional magnetic bead coating and coupling processes in existing technologies are unable to meet the requirements for MHBs detection.

[0032] To this end, the present invention significantly improves the detection performance of chemiluminescence immunoassay for MHBs mainly through hydrophobic treatment of magnetic particles and modification of MHBs antibodies.

[0033] In some embodiments of the present invention, a magnetic particle is provided, the surface of which is modified with L-valine-D8.

[0034]

[0035] In some embodiments, a method for preparing the magnetic particles is provided, comprising:

[0036] Magnetic particles are provided, and EDC and Sulfo-NHS are used to perform a first activation treatment on the magnetic particles; and then L-valine-D8 is modified on the surface of the magnetic particles.

[0037] In some embodiments, in the first activation treatment, the mass ratio of EDC to Sulfo-NHS is 1.5 to 2:1.

[0038] In some embodiments, the reaction conditions for the first activation treatment and / or the modification of L-valine-D8 include: rotating the reaction at room temperature for 30 to 120 minutes.

[0039] In some embodiments, magnetic microparticles having carboxyl groups on their surfaces may be used to perform subsequent first activation treatments, etc. For example, MS160 / Carboxyl magnetic beads may be used.

[0040] In some embodiments, the magnetic microparticles are first resuspended with an MES solution, and then EDC and Sulfo-NHS are added to the resuspended magnetic microparticles. A rotation reaction is performed at room temperature to obtain magnetic microparticles subjected to a first activation treatment. The magnetic microparticles are then magnetically separated, and the magnetic microparticles subjected to the first activation treatment are washed and resuspended with an MES solution. L-valine-D8 solution is then added, and L-valine-D8 is modified on the surface of the magnetic microparticles subjected to the first activation treatment through a rotation reaction.

[0041] In some embodiments, a magnetic particle coating is provided, comprising magnetic particles and MHBs antibodies coated on the surface of the magnetic particles; wherein,

[0042] The surface of the magnetic particles is modified with L-valine-D8; and / or,

[0043] The MHBs antibody was previously modified by reacting with the following groups: N-hydroxysuccinimide ester, pyridyl disulfide.

[0044] In some embodiments, the magnetic microparticles in the magnetic microparticle coating are surface-modified with L-valine-D8, while the MHBs antibody is not pre-modified. In other embodiments, the magnetic microparticles in the magnetic microparticle coating are not surface-modified with L-valine-D8, while the MHBs antibody is pre-modified as described above. In still other embodiments, the magnetic microparticles in the magnetic microparticle coating are surface-modified with L-valine-D8, and the MHBs antibody is pre-modified as described above. It is understood that the magnetic microparticle coating exhibits optimal detection performance when used for immunoassays when both the magnetic microparticles surface-modified with L-valine-D8 and the pre-modified MHBs antibody are used.

[0045] In some embodiments, the method for modifying the surface of the magnetic microparticles with L-valine-D8 comprises: providing magnetic microparticles, performing a first activation treatment on the magnetic microparticles with EDC and Sulfo-NHS; and then modifying the surface of the magnetic microparticles with L-valine-D8.

[0046] In some embodiments, the MHBs antibody is pre-modified by reacting with Sulfo-LC-SPDP.

[0047] In some embodiments, the MHBs antibody includes monoclonal antibody Q19 / 10, which is from Wuhan Dexin Q19 / 10.

[0048] In some embodiments, the MHBs antibody is mixed with Sulfo-LC-SPDP and incubated at 35-37° C. for 10-16 hours, thereby modifying the MHBs antibody.

[0049] In some embodiments, a method for preparing the magnetic particle coating is provided, comprising:

[0050] The magnetic particles are subjected to a second activation treatment using EDC and Sulfo-NHS to obtain activated magnetic particles; the MHBs antibody is mixed with the activated magnetic particles to perform a coating reaction to prepare the magnetic particle coating.

[0051] In some embodiments, in the second activation treatment, the mass ratio of EDC to Sulfo-NHS is 1.5 to 2:1.

[0052] In some embodiments, the magnetic microparticles are resuspended with MES solution, and then EDC and Sulfo-NHS are added, and a rotation reaction is performed at room temperature to obtain second-activated magnetic microparticles, which are then magnetically separated, and the second-activated magnetic microparticles are washed and resuspended with MES solution; then, an MHBs antibody solution is added, and the surface of the second-activated magnetic microparticles is coated with MHBs antibodies through a rotation reaction.

[0053] In some embodiments, the surface of the magnetic microparticles is modified with L-valine-D8, and the L-valine-D8 is modified by the following steps:

[0054] Magnetic particles are provided, and EDC and Sulfo-NHS are used to perform a first activation treatment on the magnetic particles; and then L-valine-D8 is modified on the surface of the magnetic particles.

[0055] In some embodiments, in the first activation treatment, the mass ratio of EDC to Sulfo-NHS is 1.5 to 2:1.

[0056] In some embodiments, the reaction conditions for the first activation treatment and / or the modification of L-valine-D8 include: rotating the reaction at room temperature for 30 to 120 minutes.

[0057] In some embodiments, the MHBs antibody is pre-modified by reacting with Sulfo-LC-SPDP.

[0058] In some embodiments, after the coating reaction, the following steps are further included: curing reaction;

[0059] The curing reaction includes adding EDC and mixing at room temperature for 30 to 60 minutes. For example, an EDC solution dissolved in pre-cooled (e.g., refrigerated at 4°C) ultrapure water can be added to a concentration of about 1 to 5 mg / mL, such as 1 gm / mL, 2 gm / mL, 3 gm / mL, 4 gm / mL, 5 gm / mL, etc.

[0060] In some embodiments, a detection kit for hepatitis B virus protein is provided, which includes the magnetic microparticle coating described in any embodiment of the present application, or the magnetic microparticle coating prepared according to the preparation method described in any embodiment of the present application.

[0061] In some embodiments, the magnetic particles, magnetic particle coatings or detection kits are used for preparing immunoassay products.

[0062] In some embodiments, the immunoassay product is used to detect proteins in hepatitis B virus.

[0063] The following examples of the present invention are provided, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these examples are for illustrating the present invention, rather than for limiting the present invention.

[0064] Example 1

[0065] This embodiment provides hydrophobically modified magnetic beads, which are mainly prepared by modifying the surface of magnetic beads with L-valine-D8. The specific steps include:

[0066] 1. Prepare the required materials and equilibrate them to room temperature;

[0067] 2. Measure 0.1 mL of MS160 / Carboxyl 1.5 μm magnetic beads (100 mg / mL, purchased from JSR LifeSciences), wash them three times with 1 mL of 20 mM MES solution (pH 6.0), and resuspend the beads in 5 mL of 20 mM MES solution (pH 6.0);

[0068] 3. Use pure water to mix Pierce TM Dissolve EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) and Sulfo-NHS (N-hydroxysulfosuccinimide) into 10 mg / mL and mix them at a ratio of 1.5:1. Add 2 mL of Pierce TM The EDC and Sulfo-NHS mixture was placed in a room temperature mixer, mixed, and rotated for 30 minutes;

[0069] 4. After the reaction is completed, perform magnetic separation, discard the supernatant, add 20 mL of 20 mM MES solution (pH 6.0) to wash twice, and resuspend the magnetic beads with 19.5 mL of pre-cooled 20 mM MES solution (pH 6.0);

[0070] 5. Dissolve L-valine-D8 (purchased from Sigma) in DMSO solution and adjust the concentration to 10 mg / mL. Add 0.5 mL of L-valine-D8 solution to the resuspended magnetic beads, mix well, and rotate for 2 hours.

[0071] 6. Wash once with 20 mL of 20 mM MES solution (pH 6.0), dilute to 10 mg / mL solution with 10 mL of 20 mM MES solution (pH 6.0), and store at 2-8 ° C for later use.

[0072] Example 2

[0073] This example modifies antibodies against the hepatitis B virus middle protein (MHBs). Specifically, the monoclonal antibody Q19 / 10, obtained from Wuhan Dexin Q19 / 10, is used. MHBs antibodies specifically recognize MHBs and do not immunoreact with LHBs and SHBs. The specific steps include:

[0074] 1. Prepare the required materials and equilibrate them to room temperature;

[0075] 2. Measure 3 mg of MHBs antibody into a reaction tube, add 0.05 mL of 1 mg / mL Sulfo-LC-SPDP (sulfosuccinimide 6-(3'-(2-pyridyldithio)propionylamino)hexanoate), and incubate in a 37°C incubator for 12 hours. After equilibration to room temperature, transfer to a 2-8°C refrigerator and incubate for 6 hours to achieve the optimal state for the reaction. After incubation, desalt and remove impurities to prepare the modified MHBs antibody.

[0076] Example 3

[0077] In this example, the hydrophobically modified magnetic beads prepared in Example 1 were coupled with the modified MHBs antibody prepared in Example 2. The specific steps included:

[0078] 1. Prepare the required materials and equilibrate them to room temperature;

[0079] 2. Wash the hydrophobically modified magnetic beads (100 mg, 10 mg / mL) three times with 1 mL of 20 mM MES solution (pH 6.0) and resuspend in 5 mL of 20 mM MES solution (pH 6.0);

[0080] 3. Use pure water to mix Pierce TM EDC and Sulfo-NHS were dissolved into 10 mg / mL and mixed at a ratio of 2:1. 2.5 mL of Pierce TM Mix the EDC and Sulfo-NHS solution, mix well, place in a mixer at room temperature, and rotate for 30 minutes;

[0081] 4. After the reaction is completed, perform magnetic separation, discard the supernatant, add 20 mL of 20 mM MES solution (pH 6.0) to wash once, and resuspend with 10 mL of 20 mM MES solution (pH 6.0);

[0082] 5. Add 3 mg of the modified MHBs antibody solution to the resuspended system in step 4, mix well, place in a room temperature mixer, and carry out coating reaction under rotating conditions for 3 hours;

[0083] 6. After the coating reaction is completed, the magnetic bead coating is obtained and solidified: add 1 mL of pre-cooled ultrapure water to dissolve it into 5 mg / mL Pierce TM EDC, placed in a room temperature mixer and mixed for 1 hour;

[0084] 7. After the solidification reaction is completed, wash with 20 mL of TBS-T (25 mM Tris-HCl pH 7.2, 0.15 M NaCl, 0.05% Tween 20) three times, mixing for 5 minutes each time in a room temperature mixer;

[0085] 8. Perform blocking treatment: Add the blocking solution to the reaction system obtained in step 7 and mix well. Place the mixture in a room temperature mixer and rotate for blocking for 5 hours to prepare magnetic beads coupled with MHBs antibodies, which are referred to as magnetic bead coating 1.

[0086] test:

[0087] Provide MHBs samples with a concentration gradient, where the concentrations of each sample are: S1, <0.50 ng / mL; S2, 3 ng / mL; S3, 10 ng / mL; S4, 40 ng / mL; S5, 100 ng / mL. Follow the steps below to perform detection using an automatic chemiluminescence analyzer:

[0088] The magnetic bead coating was diluted to 0.16 mg / mL, 50 μL of the magnetic bead coating dilution solution was taken, 100 L of the test sample solution was added and incubated at 37 ° C for 20 min, and then magnetic separation was performed to obtain the first reactant. The first reactant was washed twice with pH 7.4 PBS buffer, 50 μL of acridinium ester-labeled MHBs antibody A (from Wuhan Dexin Q19 / 10-A, which can be paired with MHBs antibody) was added, incubated at 37 ° C for 10 min, and then magnetic separation was performed to obtain the second reactant. The second reactant was washed twice with pH 7.4 PBS buffer, 100 μL of luminescent liquid was added, and the luminescent signal value was measured by a chemiluminescence analyzer. The test was repeated twice, and the average value of the luminescent signal value was calculated.

[0089] The above test was performed using freshly prepared magnetic bead coating 1 and magnetic bead coating 1 stored at 37°C for 7 days (referred to as 37°C accelerated 7 days, used to simulate the stability of 6 to 12 months under normal storage conditions), and the deviation was calculated. The results are shown in Table 1.

[0090] Table 1

[0091] sample Before acceleration processing, RLU mean 37℃ accelerated for 7 days, RLU average deviation S1 258.5 257.5 -0.4％ S2 4510 4594.5 1.9％ S3 36492.5 37766 3.5％ S4 154174 145715.5 -5.5％ S5 253813.5 273313 7.7％

[0092] As can be seen from Table 1, the use of the magnetic bead coating 1 provided in this embodiment for MHBs antigen detection has the advantages of low background and high signal-to-noise ratio, and can still meet the detection requirements after being accelerated at 37°C for 7 days.

[0093] Comparative Example 1

[0094] Magnetic beads without hydrophobic modification were coupled with unmodified MHBs antibodies without solidification treatment to prepare magnetic bead coating 2.

[0095] The preparation and testing were performed according to the same steps as in Example 3, except that MS160 / Carboxyl 1.5 μm magnetic beads and unmodified MHBs antibody were used, and step 6 was omitted. The test results are shown in Table 2.

[0096] Table 2

[0097] sample Before acceleration processing, RLU mean S1 456 S2 1505 S3 11400 S4 47550 S5 80299

[0098] The above experimental results show that when unmodified magnetic beads and MHBs antibodies are directly coupled, the magnetic bead-coated materials prepared have high background and low signal-to-noise ratio when used for MHBs antigen detection, which cannot meet the detection requirements.

[0099] Example 4

[0100] In this example, the hydrophobically modified magnetic beads prepared in Example 1 were coupled with the modified MHBs antibody prepared in Example 2, but no solidification treatment was performed, to prepare a magnetic bead coating 3.

[0101] Except for omitting step 6, the same steps as in Example 3 were followed for preparation and testing. The results are shown in Table 3.

[0102] Table 3

[0103] sample Before acceleration processing, RLU mean 37℃ accelerated for 7 days, RLU average deviation S1 249.5 312 25.1％ S2 4388.5 3489 -20.5％ S3 37992.5 33266 -12.4％ S4 167720 140715.5 -16.1％ S5 255865.5 198313 -22.5％

[0104] The above results demonstrate that the magnetic bead coating 3 provided in this example has the advantages of low background and high signal-to-noise ratio for MHBs antigen detection, meeting the basic requirements of detection. However, the results of the 37°C accelerated test indicate that its stability decreases after long-term storage, making it difficult to continue to meet detection requirements.

[0105] Example 5

[0106] In this example, the hydrophobically modified magnetic beads prepared in Example 1 were coupled with unmodified MHBs antibodies without performing a solidification treatment to prepare a magnetic bead coating 4.

[0107] The preparation and testing were carried out according to the same steps as in Example 3, except that unmodified MHBs antibody was used and step 6 was omitted. The test results are shown in Table 4.

[0108] Table 4

[0109] sample Before acceleration processing, RLU mean S1 468.5 S2 3285.5 S3 39000 S4 162550 S5 262772

[0110] The above results show that by using the magnetic bead coating 4 provided in this embodiment to detect MHBs antigen, the chemiluminescence signal is improved compared with the control example, and has a higher signal-to-noise ratio.

[0111] Example 6

[0112] The magnetic beads without hydrophobic modification were coupled with the modified MHBs antibody prepared in Example 2 without performing a solidification treatment to prepare a magnetic bead coating 5.

[0113] The preparation and testing were carried out according to the same steps as in Example 3, except that MS160 / Carboxyl 1.5 μm magnetic beads were used and step 6 was omitted. The test results are shown in Table 5.

[0114] Table 5

[0115] sample Before acceleration processing, RLU mean S1 293 S2 1892 S3 14134.5 S4 59200 S5 100957

[0116] The above results show that by using the magnetic bead coating 5 provided in this embodiment to detect MHBs antigen, the background level of the chemiluminescent reagent is reduced to the target level compared with the control example.

[0117] The foregoing description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications or substitutions that can be readily conceived by a person skilled in the art within the technical scope disclosed herein are intended to be encompassed within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be subject to the scope of protection of the claims.

Claims

1. A magnetic particle, characterized in that: The surface of the magnetic particles is modified with L-valine-D8.

2. The method for preparing magnetic particles according to claim 1, characterized in that: include: Magnetic particles are provided, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysulfosuccinimide are used to perform a first activation treatment on the magnetic particles; and then L-valine-D8 is modified on the surface of the magnetic particles.

3. The preparation method according to claim 2, wherein In the first activation treatment, the mass ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to N-hydroxysulfosuccinimide is 1.5 to 2:1; Preferably, the reaction conditions of the first activation treatment and / or the modification of L-valine-D8 include: rotating reaction at room temperature for 30 to 120 minutes.

4. A magnetic particle coating, characterized in that: It comprises magnetic particles and hepatitis B virus protein antibodies coated on the surface of the magnetic particles; wherein, The surface of the magnetic particles is modified with L-valine-D8; and / or, The hepatitis B virus middle protein antibody is modified in advance by reacting with the following groups: N-hydroxysuccinimide ester, pyridyl disulfide.

5. The magnetic particle coating according to claim 4, wherein The hepatitis B virus middle protein antibody is modified in advance by reacting with sulfosuccinimidyl 6-(3'-(2-pyridyldithio)propionylamino)hexanoate.

6. The method for preparing the magnetic particle coating according to claim 4 or 5, characterized in that: include: The magnetic particles are subjected to a second activation treatment using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysulfosuccinimide to obtain activated magnetic particles; the hepatitis B virus protein antibody is mixed with the activated magnetic particles to perform a coating reaction to prepare the magnetic particle coating.

7. The preparation method according to claim 6, wherein The surface of the magnetic particles is modified with L-valine-D8, and the L-valine-D8 is modified by the following steps: Providing magnetic particles, performing a first activation treatment on the magnetic particles using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysulfosuccinimide; and then modifying the surface of the magnetic particles with L-valine-D8; Preferably, in the first activation treatment, the mass ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to N-hydroxysulfosuccinimide is 1.5 to 2:1; Preferably, the reaction conditions of the first activation treatment and / or the modification of L-valine-D8 include: rotating reaction for 30 to 120 minutes at room temperature; Preferably, the MHBs antibody is modified in advance by reacting with Sulfo-LC-SPDP.

8. The preparation method according to claim 6 or 7, characterized in that After the coating reaction, the following steps are also included: curing reaction; The curing reaction includes adding 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and mixing at room temperature for 30 to 60 minutes.

9. A detection kit for hepatitis B virus protein, characterized in that: The invention comprises the magnetic particle coating according to claim 4 or 5, or the magnetic particle coating prepared by the preparation method according to any one of claims 6 to 8.

10. Use of the magnetic particles according to claim 1, the magnetic particle coating according to claim 4 or 5, or the detection kit according to claim 9 for preparing immunoassay products. Preferably, the immunoassay product is used to detect proteins in hepatitis B virus.

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