Detection kit for autoimmune encephalitis antibody and preparation method thereof

Abstract

The invention relates to a detection kit for an autoimmune encephalitis antibody and a preparation method of the detection kit. The detection kit for the antibody against myasthenia gravis comprises the following components: a myasthenia gravis antibody detection plate, a working solution, PBS (Phosphate Buffer Solution), a fluorescent secondary antibody and a mounting medium. According to the technical scheme, the carboxymethyl cellulose modified trehalose is used as the carrier of the antibody detection material to adsorb and wrap the cells, so that the fixing efficiency and stability of the cells on the detection carrier are remarkably improved, and the biological activity maintaining capability of the cells is enhanced, thereby improving the sensitivity and specificity of antibody detection and further improving the detection accuracy of the antibody. The carboxymethyl cellulose modified trehalose also can provide good biocompatibility, has excellent surface stability, can maintain the natural conformation and function of cells in the detection process, and effectively reduces the detection error caused by antigen denaturation or degradation.

Description

Technical Field

[0001] The invention relates to the technical field of cell detection, in particular to a detection kit for autoimmune encephalitis antibodies and a preparation method thereof. Background Art

[0002] Autoimmune encephalitis is a type of inflammatory disease of the central nervous system caused by abnormal immune system. These diseases are usually related to autoantibodies produced in the body attacking specific antigenic proteins in the nervous system. In recent years, studies have shown that a variety of antibodies (such as NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65, etc.) are closely related to the onset of autoimmune encephalitis. The detection of these antibodies is of great clinical significance for the early diagnosis, typing and formulation of treatment plans of the disease.

[0003] At present, the detection methods of autoimmune encephalitis antibodies mainly rely on laboratory immunological techniques, such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), flow cytometry, Western blotting, and immunofluorescence (IF), etc. These technologies can identify and quantify antibodies to a certain extent, but there are still some shortcomings in practical applications.

[0004] Existing immunological detection technologies often require strict experimental conditions and complex operating procedures. For example, the ELISA detection process requires multiple steps of antigen coating, washing, reaction and color development, which is time-consuming and has high requirements for experimental equipment and operating skills. Although radioimmunoassay (RIA) has high sensitivity, it involves the use of radioactive isotopes, which poses certain safety risks and environmental pollution problems. In addition, immunoblotting and flow cytometry require a high level of technology for protein separation, transfer, staining or cell processing, making it difficult to achieve large-scale screening.

[0005] In these detection methods, antigen proteins are usually fixed on solid carriers, such as glass slides, microplates or microbeads. Traditional fixation methods mainly rely on physical adsorption or chemical cross-linking, which have limitations in terms of antigen fixation efficiency and protein structure retention. Physical adsorption can easily lead to nonspecific adsorption and denaturation of antigen molecules, affecting the binding efficiency of antigens and antibodies; while chemical cross-linking methods can enhance antigen fixation, they often require the use of highly toxic cross-linking agents, which may have a negative impact on the biological activity of the antigen and reduce the sensitivity and specificity of the detection.

[0006] In addition, a major challenge in antibody detection is how to effectively maintain the stability and biological activity of antigen proteins. In traditional detection methods, antigen proteins are easily affected by environmental conditions (such as temperature, humidity, and solution composition) after being fixed on the surface of the carrier, causing denaturation or degradation, thereby affecting the accuracy of the detection. At the same time, these methods are usually difficult to achieve the coexistence and stable display of multiple antigens, and therefore it is difficult to meet the needs of simultaneous detection of multiple autoimmune encephalitis antibodies.

[0007] In order to solve these problems, new materials and technologies have been gradually introduced into the existing technology. For example, some studies have tried to use new materials with good biocompatibility and strong adsorption capacity, such as porous silicon, nanofibers or hydrogels, as antigen protein fixation carriers to improve the fixation efficiency and stability of antigen proteins. However, these materials also have some problems in application, such as complex preparation process, high material cost, nonspecific adsorption and uneven distribution of antigen proteins, etc., resulting in poor reproducibility and consistency of detection.

[0008] Therefore, it is urgent to develop a detection kit for autoimmune encephalitis antibodies to solve the above problems. Summary of the invention

[0009] Based on this, it is necessary to provide a new type of autoimmune encephalitis antibody detection kit. This material and this technical solution proposes to use carboxymethyl cellulose-modified trehalose as the detection material. Trehalose is a disaccharide substance with natural protective function, which can stabilize the natural conformation of protein in a dry environment. However, due to its high water solubility, it is easy to lose when used directly, and it is difficult to use as a fixed matrix. By blending and modifying trehalose with carboxymethyl cellulose, on the one hand, its binding force and fixation effect on antigen proteins are enhanced, and on the other hand, its surface stability and ability to resist environmental influences are improved, thereby ensuring that the structure and activity of the antigen protein are not destroyed during the detection process. In addition, multiple antigen proteins are effectively displayed on the same test plate, supporting the simultaneous detection of multiple autoimmune encephalitis antibodies.

[0010] One object of the present invention is to provide a detection kit for autoimmune encephalitis antibodies, wherein the detection kit for myasthenia gravis antibodies comprises the following components: a myasthenia gravis antibody detection plate, a working solution, PBS, a fluorescent secondary antibody, and a sealing agent;

[0011] The autoimmune encephalitis antibody detection plate is a mixture of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins and trehalose modified with carboxymethyl cellulose, and then spotted on a glass slide.

[0012] Furthermore, positive controls and negative controls are included.

[0013] Furthermore, the working fluid is goat serum or bovine serum.

[0014] Furthermore, the fluorescent secondary antibody is stained with fluorescein isothiocyanate.

[0015] Furthermore, the sealing medium is glycerol.

[0016] Furthermore, the negative control is 8-12 wt % goat serum.

[0017] Furthermore, the positive control is a recombinant antigen containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 proteins.

[0018] Specifically, autoimmune encephalitis is associated with antibodies against the surface of neuronal cells or synaptic proteins. In recent years, the spectrum of chronic inflammatory brain diseases characterized by the presence of antigen-specific antibodies in serum and cerebrospinal fluid has expanded significantly. Since there are many similarities in the clinical, imaging and laboratory manifestations of autoimmune encephalitis and infectious encephalitis, their clinical diagnosis is somewhat difficult. It has been reported that positive autoantibodies to neural antigens such as NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 are highly suggestive of the presence of autoimmune encephalitis. This product comprehensively detects NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1 and GAD65 autoantibodies, which is helpful for the auxiliary diagnosis of limbic encephalitis and encephalitis caused by unknown causes, and provides a basis for the clinical diagnosis of autoimmune encephalitis and the differential diagnosis of other diseases.

[0019] The present invention also provides a method for preparing the detection kit for the autoimmune encephalitis antibody, comprising the following steps:

[0020] S1. adding carboxylated trehalose and an activator into water and blending them, cooling and activating them to obtain an intermediate product;

[0021] S2. blending the intermediate product with a carboxymethyl cellulose solution, reacting, and purifying to obtain carboxymethyl cellulose-modified trehalose;

[0022] S3. Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 proteins are mixed with trehalose modified with carboxymethyl cellulose, spotted on a glass slide, and cut to form the test strip to obtain the antibody detection kit for autoimmune encephalitis.

[0023] Furthermore, in step S2, the heating temperature is 25-35°C.

[0024] The present invention has the following beneficial effects:

[0025] This technical solution uses carboxymethyl cellulose-modified trehalose as a carrier of antibody detection material to adsorb and wrap cells, significantly improving the fixation efficiency and stability of cells on the detection carrier, enhancing the ability to maintain the biological activity of cells, thereby improving the sensitivity and specificity of antibody detection, and carboxymethyl cellulose-modified trehalose can also provide good biocompatibility, and its surface stability is excellent, and it can maintain the natural conformation and function of cells during the detection process, effectively reducing the detection error caused by antigen denaturation or degradation. Moreover, the detection material of the present invention can realize the simultaneous detection of multiple autoimmune encephalitis-related antibodies (such as NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65). This multiple detection capability significantly improves the detection efficiency, reduces sample consumption and operation time, and meets the clinical demand for rapid detection of multiple antibodies. Through this technical solution, not only the problems of unstable antigen fixation, low detection sensitivity, and complex operation in traditional detection materials are solved, but also a more efficient and flexible antibody detection platform is provided, which can be widely used in the early diagnosis, course monitoring and treatment evaluation of autoimmune encephalitis, and provides important support for clinical practice and scientific research. DETAILED DESCRIPTION

[0026] In order to more clearly illustrate the technical solution of the present invention, the following examples are listed. Unless otherwise stated, the raw materials, reactions and post-treatment methods shown in the examples are common raw materials on the market and technical methods well known to those skilled in the art.

[0027] The words "preferred", "preferably", "more preferably", etc. in the present invention refer to embodiments of the present invention that can provide certain beneficial effects in certain circumstances. However, other embodiments may also be preferred under the same circumstances or other circumstances. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the present invention.

[0028] It should be understood that, except in any operating examples, or where otherwise indicated, all numbers expressing, for example, the amounts of ingredients used in the specification and claims should be understood to be modified in all instances by the term "about". Therefore, unless indicated to the contrary, the numerical parameters set forth in the following specification and the appended claims are approximate values ​​that vary depending on the desired properties to be obtained by the present invention.

[0029] Trehalose was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0030] Carboxymethyl cellulose, molecular weight 250000, DS=0.7.

[0031] HeLa cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins were purchased from Beijing Jingyi Medical Testing Laboratory Co., Ltd.

[0032] Example 1

[0033] A detection kit for autoimmune encephalitis antibodies, the myasthenia gravis antibody detection kit comprising the following components: the myasthenia gravis antibody detection kit comprising the following components: 2 autoimmune encephalitis antibody detection plates, 2 working solutions (10g / 100ml goat serum in PBS buffer), 20 PBS (0.01mol / L), 0.5 fluorescent secondary antibody (with fluorescein isothiocyanate (FITC)), 5 mounting media (glycerol), 2 positive controls (containing recombinant antigens of NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins) and 2 negative controls (10wt% goat serum);

[0034] The autoimmune encephalitis antibody detection plate is a mixture of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins and trehalose modified with carboxymethyl cellulose, and then spotted on a glass slide.

[0035] The preparation method of the above-mentioned autoimmune encephalitis antibody detection kit comprises the following steps:

[0036] S1-1. 1.5 parts of succinic anhydride were dissolved in 30 parts of anhydrous DMF to obtain a mixture 1;

[0037] S1-2: 2 parts of triethylamine are dissolved in 27 parts of anhydrous DMF to obtain a mixed solution 2;

[0038] S1-3. Dissolve 4 parts of trehalose in 200 parts of anhydrous DMF, mix well, heat in an oil bath at 80°C, pass nitrogen, add mixed solution 1 under stirring, mix well and then add mixed solution 2, react under sealed conditions for 12h, distill under reduced pressure, precipitate with a mixed solution of ether and acetone (7:3, v / v), centrifuge, and dry to obtain carboxylated trehalose;

[0039] S1-4. 3 parts of carboxylated trehalose, 0.5 parts of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.2 parts of 4-dimethylaminopyridine were added to 15 parts of deionized water and stirred in an ice bath for 40 min for activation, and then cooled to obtain an intermediate product;

[0040] S2-1. 5 parts of the intermediate product were blended with 150 parts of a 12 mg / mL carboxymethyl cellulose solution, reacted at 30 ° C for 24 h, and dialyzed (molecular weight 8000) to obtain carboxymethyl cellulose-modified trehalose;

[0041] S2-2 2 parts of carboxymethyl cellulose-modified trehalose were dissolved in 100 parts of pure water to obtain a mixed solution;

[0042] S3. Add 5 mg / ml PBS solution of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins to the mixed solution (ensuring that the Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins are 2wt%), after mixing, spot it on a glass slide at a temperature of 25°C and a humidity of 35% to obtain an autoimmune encephalitis antibody detection plate, cut it into detection strips, and then package it together with other components according to the above-mentioned mass fractions, and freeze it in liquid nitrogen to obtain the material for detecting autoimmune encephalitis antibodies.

[0043] Example 2

[0044] A detection kit for autoimmune encephalitis antibodies, the myasthenia gravis antibody detection kit comprising the following components: the myasthenia gravis antibody detection kit comprising the following components: 3 autoimmune encephalitis antibody detection plates, 3 working solutions (10g / 100ml goat serum in PBS buffer), 22 PBS (0.01mol / L), 1 fluorescent secondary antibody (with fluorescein isothiocyanate (FITC)), 6 mounting media (glycerol), 3 positive controls (containing recombinant antigens of NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins) and 3 negative controls (10wt% goat serum);

[0045] The autoimmune encephalitis antibody detection plate is a mixture of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins and trehalose modified with carboxymethyl cellulose, and then spotted on a glass slide.

[0046] The preparation method of the above-mentioned autoimmune encephalitis antibody detection kit comprises the following steps:

[0047] S1-1. 1.5 parts of succinic anhydride were dissolved in 30 parts of anhydrous DMF to obtain a mixture 1;

[0048] S1-2: 2 parts of triethylamine are dissolved in 27 parts of anhydrous DMF to obtain a mixed solution 2;

[0049] S1-3. Dissolve 4 parts of trehalose in 200 parts of anhydrous DMF, mix well, heat in an oil bath at 80°C, pass nitrogen, add mixed solution 1 under stirring, mix well and then add mixed solution 2, react under sealed conditions for 12h, distill under reduced pressure, precipitate with a mixed solution of ether and acetone (7:3, v / v), centrifuge, and dry to obtain carboxylated trehalose;

[0050] S1-4. 3 parts of carboxylated trehalose, 0.5 parts of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.2 parts of 4-dimethylaminopyridine were added to 15 parts of deionized water and stirred in an ice bath for 40 min for activation, and then cooled to obtain an intermediate product;

[0051] S2-1. 5 parts of the intermediate product were blended with 150 parts of a 12 mg / mL carboxymethyl cellulose solution, reacted at 30 ° C for 24 h, and dialyzed (molecular weight 8000) to obtain carboxymethyl cellulose-modified trehalose;

[0052] S2-2 2 parts of carboxymethyl cellulose-modified trehalose were dissolved in 100 parts of pure water to obtain a mixed solution;

[0053] S3. Add 5 mg / ml PBS solution of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins to the mixed solution (ensuring that the Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins are 2wt%), after mixing, spot it on a glass slide at a temperature of 25°C and a humidity of 35% to obtain an autoimmune encephalitis antibody detection plate, cut it into detection strips, and then package it together with other components according to the above-mentioned mass fractions, and freeze it in liquid nitrogen to obtain the material for detecting autoimmune encephalitis antibodies.

[0054] Example 3

[0055] A detection kit for autoimmune encephalitis antibodies, the myasthenia gravis antibody detection kit comprising the following components: the myasthenia gravis antibody detection kit comprising the following components: 3 autoimmune encephalitis antibody detection plates, 2 working solutions (10g / 100ml goat serum in PBS buffer), 25 PBS (0.01mol / L), 0.5 fluorescent secondary antibody (with fluorescein isothiocyanate (FITC)), 4 mounting media (glycerol), 2 positive controls (containing recombinant antigens of NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins) and 2 negative controls (10wt% goat serum);

[0056] The autoimmune encephalitis antibody detection plate is a mixture of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins and trehalose modified with carboxymethyl cellulose, and then spotted on a glass slide.

[0057] The preparation method of the above-mentioned autoimmune encephalitis antibody detection kit comprises the following steps:

[0058] S1-1. 1.5 parts of succinic anhydride were dissolved in 30 parts of anhydrous DMF to obtain a mixture 1;

[0059] S1-2: 2 parts of triethylamine are dissolved in 27 parts of anhydrous DMF to obtain a mixed solution 2;

[0060] S1-3. Dissolve 4 parts of trehalose in 200 parts of anhydrous DMF, mix well, heat in an oil bath at 80°C, pass nitrogen, add mixed solution 1 under stirring, mix well and then add mixed solution 2, react under sealed conditions for 12h, distill under reduced pressure, precipitate with a mixed solution of ether and acetone (7:3, v / v), centrifuge, and dry to obtain carboxylated trehalose;

[0061] S1-4. 3 parts of carboxylated trehalose, 0.5 parts of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.2 parts of 4-dimethylaminopyridine were added to 15 parts of deionized water and stirred in an ice bath for 40 min for activation, and then cooled to obtain an intermediate product;

[0062] S2-1. 5 parts of the intermediate product were blended with 150 parts of a 12 mg / mL carboxymethyl cellulose solution, reacted at 30 ° C for 24 h, and dialyzed (molecular weight 8000) to obtain carboxymethyl cellulose-modified trehalose;

[0063] S2-2 2 parts of carboxymethyl cellulose-modified trehalose were dissolved in 100 parts of pure water to obtain a mixed solution;

[0064] S3. Add 5 mg / ml PBS solution of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins to the mixed solution (ensuring that the Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 proteins are 2wt%), after mixing, spot it on a glass slide at a temperature of 25°C and a humidity of 35% to obtain an autoimmune encephalitis antibody detection plate, cut it into detection strips, and then package it together with other components according to the above-mentioned mass fractions, and freeze it in liquid nitrogen to obtain the material for detecting autoimmune encephalitis antibodies.

[0065] Comparative Example 1

[0066] A detection kit for autoimmune encephalitis antibodies. The difference between this comparative example and Example 1 is that this comparative example uses trehalose of equal mass to replace trehalose modified with carboxymethyl cellulose, and other components and preparation methods are the same.

[0067] Test Case

[0068] The materials for detecting autoimmune encephalitis antibodies prepared in Example 1 and Comparative Example 1 were used as a kit for performance testing.

[0069] The materials for detecting autoimmune encephalitis antibodies prepared in Example 1 and Comparative Example 1 were used as kits to detect 20 NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 negative sera and 20 NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, GAD65 positive sera;

[0070] Test method:

[0071] A1. Equilibrate room temperature: Take out the cell matrix slide and place it at room temperature for 30 minutes;

[0072] A2. Sample incubation: Add 100 μl of the diluted serum sample to be tested to each well of the reaction plate in a fume hood, place the cell matrix slide upside down on the reaction plate, and incubate at room temperature for 1 hour;

[0073] A3. Washing: Immerse the cell matrix slide in a washing tank filled with PBS for 5 minutes, take out the cell matrix slide, and wipe off the excess liquid on the edge and back with absorbent paper;

[0074] A4. Secondary antibody incubation: Add 100 μl of fluorescent secondary antibody to each well of a dry and clean reaction plate, quickly invert the cell matrix slide onto the reaction plate, and incubate at room temperature for 1 hour;

[0075] A5. Washing: Immerse the cell matrix slide in a washing tank filled with PBS for 5 minutes in a dark environment. Take out the cell matrix slide and wipe off the excess liquid on the edge and back with absorbent paper.

[0076] A6. Sealing: Add 100 μl of sealing medium to each reaction area and cover with a coverslip;

[0077] A7. Observe the fluorescence signal: Read the green fluorescence result under the microscope, observe with a 20x objective lens, and analyze the result.

[0078] Table 1 Material performance test results for detecting autoimmune encephalitis antibodies in Example 1 and Comparative Example 1

[0079]

[0080] It can be concluded from Table 1 that the detection material of Example 1 of the present invention has high detection sensitivity and can simultaneously identify and quantify multiple autoimmune encephalitis antibodies in one detection process, thereby improving detection efficiency.

[0081] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0082] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A detection kit for autoimmune encephalitis antibodies, characterized in that: The myasthenia gravis antibody detection kit comprises the following components: a myasthenia gravis antibody detection plate, a working solution, PBS, a fluorescent secondary antibody, and a sealing agent; The autoimmune encephalitis antibody detection plate is a mixture of Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 antigen proteins and trehalose modified with carboxymethyl cellulose, and then spotted on a glass slide.

2. The antibody detection kit for autoimmune encephalitis according to claim 1, characterized in that: Positive and negative controls were also included.

3. The antibody detection kit for autoimmune encephalitis according to claim 1, characterized in that: The working solution is goat serum or bovine serum.

4. The antibody detection kit for autoimmune encephalitis according to claim 1, characterized in that: The fluorescent secondary antibody is stained with fluorescein isothiocyanate.

5. The antibody detection kit for autoimmune encephalitis according to claim 1, characterized in that: The mounting medium is glycerol.

6. The antibody detection kit for autoimmune encephalitis according to claim 2, characterized in that: The negative control is 8-12 wt % goat serum.

7. The antibody detection kit for autoimmune encephalitis according to claim 2, characterized in that: The positive control is a recombinant antigen containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 proteins.

8. A method for preparing a detection kit for autoimmune encephalitis antibodies according to any one of claims 1 to 7, characterized in that: The steps include: S1. adding carboxylated trehalose and an activator into water and blending them, cooling and activating them to obtain an intermediate product; S2. blending the intermediate product with a carboxymethyl cellulose solution, reacting, and purifying to obtain carboxymethyl cellulose-modified trehalose; S3. Hela cells containing NMDAR, GABABR, AMPAR1, AMPAR2, CASPR2, IgLON5, LGI1, and GAD65 proteins are mixed with trehalose modified with carboxymethyl cellulose, spotted on a glass slide, and cut to form the test strip to obtain the antibody detection kit for autoimmune encephalitis.

9. The method for preparing a detection kit for autoimmune encephalitis antibodies according to claim 8, characterized in that: In step S2, the heating temperature is 25-35°C.

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