Rheumatoid factor blocker, preparation method thereof, and immunoassay kit

Abstract

The present invention relates to the field of in vitro diagnostic technology, and more particularly to a rheumatoid factor blocker, a preparation method thereof, and an immunoassay kit. The preparation method comprises the following steps: dissolving a compound of formula I in a first solvent to prepare a first solution; dissolving a compound of formula II in a second solvent to prepare a second solution; mixing the first and second solutions, reacting the compound of formula I with the compound of formula II, and collecting the reaction product. The preparation method of the present invention can produce a rheumatoid factor blocker capable of reducing RF interference with immunoassays.

Description

Technical Field

[0001] The present invention relates to the technical field of in vitro diagnosis, and in particular to a rheumatoid factor blocker, a preparation method thereof, and an immunoassay kit. Background Art

[0002] Rheumatoid factor (RF) is a common immune interfering substance. It is a common clinical autoantibody that appears not only in rheumatoid arthritis but also in other autoimmune diseases (such as SLE), infectious diseases, and even in healthy individuals. RF interferes with many immunoassays, resulting in false positive or false negative results. The extent of RF interference and its concentration-dose effect are unclear; both high and low concentrations may interfere with immunoassays. Summary of the Invention

[0003] Based on this, the present invention provides a rheumatoid factor blocker, a preparation method thereof, and an immunoassay kit to reduce the interference of RF on immunoassay.

[0004] The first aspect of the present invention provides a method for preparing a rheumatoid factor blocker, and the technical solution thereof is as follows:

[0005] A method for preparing a rheumatoid factor blocker comprises the following steps:

[0006] dissolving the compound of formula I in a first solvent to prepare a first solution;

[0007] dissolving the compound of formula II in a second solvent to prepare a second solution;

[0008] mixing the first solution and the second solution, allowing the compound of formula I to react with the compound of formula II, and collecting the reaction product;

[0009]

[0010] In some embodiments, n is any integer between 1 and 24.

[0011] In some embodiments, the molar ratio of the compound of formula I to the compound of formula II is (4-32):1.

[0012] In some embodiments, the concentration of the compound of formula I in the first solution is 490 nM to 510 nM.

[0013] In some embodiments, the first solvent is selected from one of dimethyl sulfoxide and N,N-dimethylformamide.

[0014] In some embodiments, the concentration of the compound of formula II in the second solution is 28 mM to 32 mM.

[0015] In some embodiments, the second solvent is selected from one or more of phosphate buffer and carbonate buffer.

[0016] In some embodiments, the reaction of the compound of formula I with the compound of formula II is carried out under at least one of the following parameters: (1) the reaction temperature is 10° C. to 30° C.; and (2) the reaction time is 50 min to 70 min.

[0017] In some embodiments, after reacting the compound of formula I with the compound of formula II, the method further comprises adding lysine to terminate the reaction.

[0018] The second aspect of the present invention provides a rheumatoid factor blocker, which is prepared by the above preparation method.

[0019] The third aspect of the present invention provides an immunoassay kit, the technical solution of which is as follows:

[0020] An immunoassay kit comprises the above-mentioned rheumatoid factor blocker.

[0021] In some embodiments, the immunoassay kit further includes one or more of a solid phase coating and a composite marker; wherein the solid phase coating is coated with a first binding protein, the composite marker includes a coupled second binding protein and a luminescent marker, and both the first binding protein and the second binding protein can specifically bind to the immunoassay target and have different binding sites.

[0022] In some embodiments, the solid phase in the solid phase coating is magnetic beads.

[0023] In some embodiments, the luminescent label is a chemiluminescent label.

[0024] In some embodiments, when the immunodetection target is β2IgM, the first binding protein is β2 glycoprotein I, and the second binding protein is mouse anti-human IgM antibody.

[0025] In some embodiments, when the immunodetection target is IgM, the first binding protein is a recombinant rubella virus protein, and the second binding protein is a mouse anti-human IgM antibody.

[0026] In some embodiments, the immunoassay kit further comprises a sample diluent.

[0027] Compared with the traditional solution, the present invention has the following beneficial effects:

[0028] In the present invention, the compound of formula II is aspartame, and the compound of formula I is used to modify aspartame, and carboxyl polyethylene glycol is covalently linked to the aspartic acid amino terminus of aspartame through succinimide to synthesize carboxyl polyethylene glycol aspartame as an RF blocker. The RF blocker newly synthesized by the above method can enter the active site of IgM RF and bind thereto, and can inhibit its binding interaction with the human IgG Fc region, thereby reducing the interference caused by RF in the immune response. At the same time, due to the high hydrophilicity of the introduced new group, the solubility of the RF blocker in the aqueous solution can reach more than 700nM, which can make the concentration of the active ingredient in the immune response system high, and have a good blocking effect on RF. It has a good inhibitory effect on the interference of RF in the immune response, thereby improving the accuracy of immune response detection, and at the same time, does not affect the detection sensitivity. DETAILED DESCRIPTION

[0029] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to provide a more thorough and comprehensive understanding of the present disclosure.

[0030] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0031] the term

[0032] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0033] In the present invention, the selection range of "and / or", "or / and", and "and / or" includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, and the said any and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that the technical solution undoubtedly includes technical solutions connected by "logical and" and technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0034] In the present invention, "plurality", "multiple", "multiple times", "multiple", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0035] In the present invention, "combinations thereof", "any combination thereof", "any combination thereof" and the like include all suitable combinations of any two or more of the listed items.

[0036] In the present invention, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc. shall be based on the ability to implement the technical solution of the present invention, solve the technical problem of the present invention, and achieve the expected technical effect of the present invention.

[0037] In the present invention, “preferred”, “better”, “more preferred” and “suitable” are only used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute a limitation on the scope of protection of the present invention.

[0038] In the present invention, the terms “further”, “further”, “particularly”, etc. are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of the present invention.

[0039] In this disclosure, the terms "optionally," "optional," and "optional" are optional and refer to either option, i.e., to the selection of either option from the two parallel options of "optional" or "optional." If multiple "optional" terms appear in a technical solution, each "optional" term is independent unless otherwise specified and there are no conflicts or constraints.

[0040] In the present invention, in the "first aspect," "second aspect," "third aspect," "fourth aspect," etc., the terms "first," "second," "third," "fourth," etc. are used for descriptive purposes only and should not be understood as indicating or implying relative importance or quantity, nor as implicitly indicating the importance or quantity of the technical features indicated. Furthermore, "first," "second," "third," "fourth," etc. serve only as non-exhaustive enumeration and description and should be understood not to constitute a closed-ended limitation on quantity.

[0041] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0042] In the present invention, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval and includes the two numerical endpoints of the numerical range (i.e., the minimum value and the maximum value), as well as each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval refers only to integers within the numerical interval, it includes the two endpoint integers of the numerical range, as well as each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0043] The temperature parameters in the present invention, unless otherwise specified, may be either constant temperature or fluctuating within a certain temperature range. It should be understood that the constant temperature treatment allows for temperature fluctuations within the precision range of the instrument control. Fluctuations within ranges such as ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are permitted.

[0044] In the present invention, when referring to percentage concentration, unless otherwise specified, it refers to the final concentration. The final concentration refers to the percentage of the added component in the system after the addition of the component.

[0045] In the present invention, β2IgM refers to β2 glycoprotein 1 IgM type antibody, which is mainly used for auxiliary diagnosis of patients with phospholipid syndrome.

[0046] In the present invention, RF refers to rheumatoid factor, an autoantibody in the human body that can bind to human denatured IgG, mainly of the IgM type.

[0047] To address the issue of how to reduce RF interference in immunoassays, the present invention provides a method for preparing a rheumatoid factor blocker, which, in one embodiment, comprises the following steps:

[0048] dissolving the compound of formula I in a first solvent to prepare a first solution;

[0049] dissolving the compound of formula II in a second solvent to prepare a second solution;

[0050] mixing the first solution and the second solution, allowing the compound of formula I to react with the compound of formula II, and collecting the reaction product;

[0051]

[0052] In this embodiment, the compound of formula II is aspartame, which is an aspartic acid / phenylalanine dipeptide methyl ester. Studies have found that aspartame can inhibit the binding interaction with the human IgG Fc region by entering the active site of IgM RF and binding thereto, thereby reducing the interference caused by RF in the immune response. However, the solubility of aspartame in aqueous solution is only 32mM to 35mM, and it cannot completely compete for the active site of IgM RF in the immune response system. A large part of the free RF can bind to the human IgG Fc region and interfere with the immune response. If the RF in the sample is reduced by dilution, so as to use aspartame with lower solubility to reduce the interference of RF on immunoassay, the concentration of the target substance to be measured is also reduced after dilution, which will affect the sensitivity of the detection.

[0053] The reaction scheme of the preparation method of this embodiment is as follows:

[0054]

[0055] In this embodiment, aspartame is modified using a compound of formula I, and carboxyl polyethylene glycol is covalently linked to the aspartic acid amino terminus of aspartame via succinimide to synthesize carboxyl polyethylene glycol aspartame as an RF blocker. The newly synthesized RF blocker by the above method can retain the binding effect of aspartame on the activity of RF, that is, the RF blocking active site and the blocking effect remain unchanged. At the same time, because the introduced new group is highly hydrophilic, the solubility of the RF blocker in the aqueous solution can reach more than 700mM, which can increase the concentration of the active ingredient in the immune response system, improve the blocking effect on RF, and have a good inhibitory effect on the interference of RF in the immune response, thereby improving the accuracy of the immune response-based detection without affecting the detection sensitivity.

[0056] Compared with aspartame, the solubility of the RF blocker synthesized in this embodiment can be increased by more than 20 times, and the inhibitory effect on RF can be increased by more than 10 times.

[0057] Optionally, n is any integer from 1 to 24. Preferably, n is 12.

[0058] Optionally, the concentration of the compound of formula I in the first solution is 490 nM to 510 nM. It is understood that the concentration of the compound of formula I in the first solution includes but is not limited to: 490 nM, 500 nM, and 510 nM.

[0059] Optionally, the first solvent is selected from one of dimethyl sulfoxide (DMSO) and dimethylformamide (DMF).

[0060] Optionally, the concentration of the compound of formula II in the second solution is 28 mM to 32 mM. It is understood that the concentration of the compound of formula II in the second solution includes but is not limited to: 28 mM, 30 mM, 32 mM.

[0061] Optionally, the second solvent is selected from one or more of phosphate buffered saline (PB buffer) and carbonate buffered saline (CBS).

[0062] In some examples, the concentration of the phosphate buffer may be 40 mM to 60 mM. It is understood that the concentration of the phosphate buffer includes, but is not limited to, 40 mM, 50 mM, and 60 mM. Preferably, the concentration of the phosphate buffer is 50 mM.

[0063] In some examples, the pH value of the phosphate buffer can be 6.5 to 8.5. It is understood that the pH value of the phosphate buffer includes, but is not limited to, 6.5, 7.0, 7.5, 8.0, and 8.5. Preferably, the pH value of the phosphate buffer is 7.5 ± 0.1. More preferably, the pH value of the phosphate buffer is 7.5.

[0064] Optionally, before reacting the compound of Formula I with the compound of Formula II, the molar ratio of the compound of Formula I to the compound of Formula II is controlled to be (4-32):1. It is understood that the molar ratio of the compound of Formula I to the compound of Formula II includes, but is not limited to, 4:1, 8:1, 16:1, and 32:1. Preferably, the molar ratio of the compound of Formula I to the compound of Formula II is 4:1. In this case, the compound of Formula I is in excess.

[0065] In this embodiment, the compound of formula I is reacted with the compound of formula II, and the carboxyl polyethylene glycol is covalently linked to the aspartic acid amino end of aspartame via succinimide.

[0066] Optionally, the reaction of the compound of formula I with the compound of formula II is carried out under at least one of the following parameters: (1) the reaction temperature is 10° C. to 30° C.; and (2) the reaction time is 50 min to 70 min.

[0067] It can be understood that the reaction temperature includes but is not limited to: 10°C, 15°C, 20°C, 25°C, 30°C.

[0068] It can be understood that the reaction time includes but is not limited to: 50 min, 60 min, 70 min.

[0069] Optionally, after reacting the compound of formula I with the compound of formula II, the method further comprises adding lysine to terminate the reaction.

[0070] It can be understood that after the compound of formula I and the compound of formula II are fully reacted, lysine is added, and lysine reacts with the excess compound of formula I to block the active ester of the compound of formula I and terminate the reaction.

[0071] Alternatively, lysine may be added by adding a lysine aqueous solution to the reaction system, wherein the concentration of the lysine aqueous solution may be 80 mM to 120 mM.

[0072] In this embodiment, after adding lysine, the mixture is placed at 10° C. to 30° C. for 20 to 40 minutes, and the solid reaction product in the reaction solution is filtered out and freeze-dried to obtain the rheumatoid factor blocker.

[0073] It can be understood that the rheumatoid factor blocker is freeze-dried under a vacuum environment and stored at 2°C to 8°C.

[0074] The present invention also provides a rheumatoid factor blocker, which, in one embodiment, is prepared by the above-mentioned preparation method. The rheumatoid factor blocker of this embodiment can be dissolved in water before use, and its concentration in water can reach 700 nM or more, which is a high concentration and has a good blocking effect on RF.

[0075] The present invention also provides an immunoassay kit. In one embodiment, the immunoassay kit comprises the above-mentioned rheumatoid factor blocker.

[0076] Optionally, the immunoassay kit further includes one or more of a solid phase coating and a composite marker; wherein the solid phase coating is coated with a first binding protein, the composite marker includes a coupled second binding protein and a luminescent marker, and both the first binding protein and the second binding protein can specifically bind to the immunoassay target and have different binding sites.

[0077] Further optionally, the solid phase in the solid phase coating is magnetic beads.

[0078] Further optionally, the luminescent marker is a chemiluminescent marker.

[0079] Optionally, when the immunodetection target is β2IgM, the first binding protein is β2 glycoprotein I, and the second binding protein is mouse anti-human IgM antibody.

[0080] Optionally, the first binding protein is a recombinant rubella virus protein, and the second binding protein is a mouse anti-human IgM antibody.

[0081] Optionally, the immunoassay kit further comprises a sample diluent.

[0082] The following is further described in conjunction with specific examples. Unless otherwise specified, the raw materials involved in the following specific examples can all be sourced from commercial sources. The instruments used can all be sourced from commercial sources unless otherwise specified. The processes involved can all be selected by those skilled in the art unless otherwise specified.

[0083] Example 1

[0084] This embodiment provides an RF blocking agent and a preparation method thereof, and the steps are as follows:

[0085] Step 1: Dissolve the compound of formula I in DMSO to prepare a first solution with a concentration of 500 mM of the compound of formula I, and store it at -20°C in the dark, wherein n is 12 in the compound of formula I.

[0086] Step 2: dissolving the compound of formula II (aspartame) in 50 mM PB buffer to prepare a second solution with an aspartame concentration of 30 mM.

[0087] Step 3: According to the molar ratio of the compound of formula I to the compound of formula II of 4:1, the first solution and the second solution were mixed, and the reaction was carried out at 25°C for 60 minutes. A 100 mM lysine aqueous solution was added and the mixture was placed at 25°C for 30 minutes to block the active ester of the compound of formula I and terminate the reaction.

[0088] Step 4: Filter out the solid reaction product in the reaction solution after the reaction is completed, and vacuum freeze-dry to obtain carboxy polyethylene glycol aspartame, which is the RF blocker.

[0089] Example 2

[0090] The carboxyl polyethylene glycol aspartame obtained in Example 1 was dissolved in deionized water to prepare carboxyl polyethylene glycol aspartame aqueous solutions of different concentrations. The RF blocking effects of the carboxyl polyethylene glycol aspartame aqueous solutions of different concentrations were investigated.

[0091] Investigation project 1: Study on the anti-RF interference of RF blockers in β2IgM project

[0092] 1. Coat β2 glycoprotein I onto magnetic beads and prepare a magnetic bead coating working solution;

[0093] 2. Couple the acridinium ester to mouse anti-human IgM antibody and prepare an acridinium label working solution;

[0094] 3. Control group: Dispense the magnetic bead coating working solution, acridine marker working solution, and sample diluent into reagent boats to assemble the β2IgM detection reagent;

[0095] 4. In experimental group 1, 30 mM aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then dispensed into the reagent boat to assemble the β2IgM detection reagent.

[0096] 5. In experimental group 2, 30 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the solution was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then dispensed into the reagent boat to assemble the β2IgM detection reagent;

[0097] 6. In experimental group 3, 100 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then dispensed into the reagent boat to assemble the β2IgM detection reagent.

[0098] 7. In experimental group 4, 300 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then dispensed into the reagent boat to assemble the β2IgM detection reagent.

[0099] 8. Refer to the reaction parameters of the β2IgM project and test 10 RF-positive samples that can cause false positives for this project (RF interference samples 1 to 10, β2IgM has been confirmed to be negative by comparison with the manufacturer), compare the luminescence value levels under different conditions, and analyze the effects of different RF blockers and doses on improving the ability to resist RF interference.

[0100] Clinical Research:

[0101] By setting up a control group (no RF blocker added), an aspartame group (experimental group 1), and different concentrations of carboxy polyethylene glycol aspartame (experimental groups 2, 3, and 4), the anti-interference effect of carboxy polyethylene glycol aspartame and the influence of its dosage on the anti-interference ability in the β2IgM project were studied. The results are shown in Table 1. The experiment found that 30mM aspartame and 30mM carboxy polyethylene glycol aspartame can both improve the anti-interference ability of β2IgM, and can reduce the signal of the interference sample by about 30%, and some samples can be reduced by more than 50%. The anti-RF interference effects of the two are basically the same; when the concentration of carboxy polyethylene glycol aspartame is increased to 100mM and 300mM, the signal of the RF interference sample is significantly reduced, and the degree of reduction is positively correlated with the carboxy polyethylene glycol aspartame; when the concentration of carboxy polyethylene glycol aspartame is increased to 300mM, the interference signal is reduced by about 90%, and the signals of 9 out of 10 samples can be reduced to below the cutoff value, and the interference sample results are negative.

[0102] Table 1

[0103]

[0104] Investigation Project 2: Study on the Anti-RF Interference of RF Blockers in Rubella IgM (RV IgM) Project

[0105] 1. Coat the purified recombinant rubella virus protein onto magnetic beads and prepare a magnetic bead coating working solution;

[0106] 2. Couple the acridinium ester to mouse anti-human IgM antibody and prepare an acridinium label working solution;

[0107] 3. Control group: Dispense the magnetic bead coating working solution, acridine marker working solution, and sample diluent into reagent boats to assemble the RV IgM detection reagent;

[0108] 4. In experimental group 1, 30 mM aspartame aqueous solution was added to the sample diluent, and the mixture was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then dispensed into reagent boats to assemble the RV IgM detection reagent.

[0109] 5. In experimental group 2, 30 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and the mixture was dispensed into reagent boats to assemble the RV IgM detection reagent.

[0110] 6. In experimental group 3, 100 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then divided into reagent boats to assemble the RV IgM detection reagent.

[0111] 7. In experimental group 4, 300 mM carboxy polyethylene glycol aspartame aqueous solution was added to the sample diluent, and the sample was mixed with the magnetic bead coating working solution and the acridine marker working solution, and then divided into reagent boats to assemble the RV IgM detection reagent.

[0112] 8. Referring to the reaction parameters of the RV IgM project, test 10 RF-positive samples that can cause false positives for this project (RF interference samples 11 to 20, RV IgM has been confirmed to be negative by comparison with the manufacturer), compare the luminescence value levels under different conditions, and analyze the effects of different RF blockers and doses on improving the ability to resist RF interference.

[0113] Clinical Research:

[0114] A control group (no RF blocker), an aspartame group (experimental group 1), and different concentrations of carboxy-polyethylene glycol aspartame (experimental groups 2, 3, and 4) were established to investigate the anti-interference effect of carboxy-polyethylene glycol aspartame on RV IgM and the effect of its dosage on anti-interference ability. The results are shown in Table 2. The experiment found that both 30mM aspartame and 30mM carboxy polyethylene glycol aspartame can improve the anti-interference ability of RV IgM, and can reduce the signal of the interference sample by about 30%, and some samples can be reduced by more than 35%. The anti-RF interference effects of the two are basically the same; when the concentration of carboxy polyethylene glycol aspartame is increased to 100mM and 300mM, the signal of the RF interference sample is significantly reduced, and the degree of reduction is positively correlated with the carboxy polyethylene glycol aspartame; when the concentration of carboxy polyethylene glycol aspartame is increased to 300mM, the interference signal is reduced by about 90%, and the signals of 10 out of 10 samples can be reduced to below the cutoff value, and the results of the interference samples are all negative.

[0115] Table 2

[0116]

[0117]

[0118] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0119] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A method for preparing a rheumatoid factor blocker, characterized in that: The following steps are included dissolving the compound of formula I in a first solvent to prepare a first solution; dissolving the compound of formula II in a second solvent to prepare a second solution; mixing the first solution and the second solution, allowing the compound of formula I to react with the compound of formula II, and collecting the reaction product; The molar ratio of the compound of formula I to the compound of formula II is (4-32):

1.

2. The method for preparing the rheumatoid factor blocker according to claim 1, characterized in that: The n is any integer from 1 to 24.

3. The method for preparing the rheumatoid factor blocking agent according to claim 2, wherein: The concentration of the compound of formula I in the first solution is 490nM to 510nM; and / or The first solvent is selected from one of dimethyl sulfoxide and N,N-dimethylformamide; and / or The concentration of the compound of formula II in the second solution is 28mM to 32mM; and / or The second solvent is selected from one or more of a phosphate buffer and a carbonate buffer.

4. The method for preparing the rheumatoid factor blocking agent according to any one of claims 1 to 3, characterized in that: The parameters for reacting the compound of formula I with the compound of formula II are at least one of the following conditions: (1) the reaction temperature is 10° C. to 30° C.; and (2) the reaction time is 50 min to 70 min.

5. The method for preparing the rheumatoid factor blocking agent according to any one of claims 1 to 3, characterized in that: After the compound of formula I reacts with the compound of formula II, the method further comprises adding lysine to terminate the reaction.

6. A rheumatoid factor blocker, characterized in that: Prepared by the preparation method according to any one of claims 1 to 5.

7. An immunoassay kit, characterized in that: Including the rheumatoid factor blocker according to claim 6.

8. The immunoassay kit according to claim 7, characterized in that The immunoassay kit further includes one or more of a solid phase coating and a composite marker; wherein the solid phase coating is coated with a first binding protein, and the composite marker includes a coupled second binding protein and a luminescent marker, and both the first binding protein and the second binding protein can specifically bind to the immunoassay target and have different binding sites.

9. The immunoassay kit according to claim 8, characterized in that The immunoassay kit further comprises a sample diluent.

10. The immunoassay kit according to claim 9, characterized in that Under the condition that the immunodetection target is β2IgM, the first binding protein is β2 glycoprotein I, and the second binding protein is mouse anti-human IgM antibody; or, Under the condition that the immune detection target is IgM, the first binding protein is a recombinant rubella virus protein, and the second binding protein is a mouse anti-human IgM antibody.

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