Monoclonal antibody 1G6-60B12 for resisting p-Tau217, product based on monoclonal antibody 1G6-60B12 and application of monoclonal antibody 1G6-60B12

Abstract

The invention discloses a monoclonal antibody 1G6-60B12 resisting p-Tau217, a product based on the monoclonal antibody 1G6-60B12 and application of the monoclonal antibody 1G6-60B12, and belongs to the technical field of monoclonal antibodies. The invention discloses an amino acid sequence of a variable region complementary determining region of a light chain and a heavy chain of an anti-p-Tau217 monoclonal antibody 1G6-60B12, and experiments prove that the anti-p-Tau217 monoclonal antibody 1G6-60B12 has relatively high binding force, can specifically recognize a target antigen p-Tau217 oligopeptide, and can be used for preparing the anti-p-Tau217 monoclonal antibody 1G6-60B12. The compound can be used for preparing a product for detecting the p-Tau217 antigen level or preparing a product for diagnosing diseases related to the p-Tau217 antigen, and the diseases are neurodegenerative diseases such as Alzheimer's disease.

Description

Technical Field

[0001] The present invention belongs to the technical field of monoclonal antibodies, and specifically relates to an anti-p-Tau217 monoclonal antibody, as well as products and applications based on the antibody. Background Art

[0002] Alzheimer's disease (AD), a common and devastating neurodegenerative disorder, has placed a heavy burden on the world. The main pathological features of AD include the aggregation of β-amyloid protein (Aβ) outside neurons to form senile plaques, the hyperphosphorylation of tau protein within neurons to form neurofibrillary tangles (NFTs), brain atrophy caused by neuronal loss, synaptic damage, and glial cell proliferation. Massive neuronal death and loss are the core factors that lead to severe brain damage in patients.

[0003] In recent years, researchers have discovered that partially phosphorylated tau (p-tau) forms, such as p-tau181 and p-Tau217, increase significantly in the early stages of AD and can serve as highly promising biomarkers for the early diagnosis of AD. Among them, the phosphorylation of the p-Tau217 site is particularly noteworthy. Studies have shown that in patients with autosomal dominant AD, the level of p-Tau217 shows a significant upward trend, which gives it a significant advantage over other tau phosphorylation sites in distinguishing AD patients from non-AD patients (Moon H, Chen X, The journal of prevention of Alzheimer's disease. 2025: 100252). At the same time, the level of p-Tau217 in plasma begins to increase in the early stages of AD. This feature makes it an important biomarker to assist in AD longitudinal research, drug development, and disease diagnosis, monitoring and patient care (Malotaux V, Ashton NJ, Alzheimer's & dementia: the journal of the Alzheimer's Association. 2025, 21(7): e70421). Summary of the Invention

[0004] The purpose of the present invention is to provide an anti-p-Tau217 monoclonal antibody with high binding affinity that can specifically recognize the target antigen p-Tau217, and to provide products and applications based on the antibody, so as to solve the defects of reagents / kits for detecting the p-Tau217 antigen in the prior art, such as insufficient sensitivity or low specificity.

[0005] To achieve the above objectives, the present invention provides an anti-p-Tau217 monoclonal antibody (named 1G6-60B12) comprising a light chain and a heavy chain, the light chain is of kappa type and the heavy chain is of IgG1 type; wherein: the amino acid sequences of the three complementary determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region are shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3, respectively; the amino acid sequences of the three complementary determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region are shown in SEQ ID No. 9, SEQ ID No. 10 and SEQ ID No. 11, respectively.

[0006] Furthermore, the framework regions LFR1, LFR2, LFR3 and LFR4 of the light chain variable region respectively have homologous sequences with at least 90% sequence identity to the amino acid sequences shown in SEQ ID No.4, SEQ ID No.5, SEQ ID No.6 and SEQ ID No.7; the framework regions HFR1, HFR2, HFR3 and HFR4 of the heavy chain variable region respectively have homologous sequences with at least 90% sequence identity to the amino acid sequences shown in SEQ ID No.12, SEQ ID No.13, SEQ ID No.14 and SEQ ID No.15.

[0007] Furthermore, the light chain variable region VL of the monoclonal antibody 1G6-60B12 has a homologous sequence with at least 90% sequence identity to the amino acid sequence shown in SEQ ID No. 8; the heavy chain variable region VH has a homologous sequence with at least 90% sequence identity to the amino acid sequence shown in SEQ ID No. 16.

[0008] The present invention further provides use of the anti-p-Tau217 monoclonal antibody 1G6-60B12 in preparing a product for detecting the level of the p-Tau217 antigen or preparing a product for diagnosing a disease associated with the p-Tau217 antigen, wherein the disease is a neurodegenerative disease, such as Alzheimer's disease.

[0009] The present invention provides a nucleic acid molecule encoding the above-mentioned anti-p-Tau217 monoclonal antibody 1G6-60B12 or a functional fragment based thereon.

[0010] The present invention provides a recombinant expression vector containing the above nucleic acid molecule.

[0011] The present invention provides a host cell, which contains the above-mentioned recombinant expression vector.

[0012] The present invention further provides a product for detecting the level of p-Tau217 antigen, which comprises the above-mentioned nucleic acid molecule, recombinant expression vector and host cell.

[0013] Furthermore, in the product for detecting the level of p-Tau217 antigen, the recombinant expression vector has a signal peptide connected to the antibody and contains a transcriptional regulatory element; the host cell is selected from mammalian cells, specifically 293T cells, CHO cells, Expi293F cells, TM cell.

[0014] The above-mentioned product for detecting the p-Tau217 antigen level is a detection reagent, a detection kit or a test paper, wherein the detection reagent is a reagent for performing an antigen-antibody reaction or a reagent for a detection reaction; the reagent for performing an antigen-antibody reaction includes a buffer (PBS, etc.), a salt (NaCl, etc.), and a diluent (Binding buffer, Elution buffer, etc.); the kit is any one of a colloidal gold immunoassay kit, a chemiluminescence detection kit, a radioimmunoassay kit, an enzyme-linked immunosorbent assay kit, a fluorescence immunoassay kit, and a microfluidic chip.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] The anti-p-Tau217 monoclonal antibody 1G6-60B12 provided by the present invention has high binding affinity and can specifically recognize the target antigen p-Tau217 short peptide. It can be used to prepare products for detecting p-Tau217 antigen levels and preparing products for diagnosing diseases related to the p-Tau217 antigen. It is conducive to the discovery and screening of patients with early Alzheimer's disease and has important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the titer measurement result of tail tip blood of mice immunized with p-Tau217 short peptide.

[0018] Figure 2 This is the subtype identification result of monoclonal antibody 1G6-60B12.

[0019] Figure 3 This is the electrophoresis result of monoclonal antibody 1G6-60B12; M is Marker; 1 is ascites; 2 is flow-through; 3 is Elution Buffer; 4 is ultrafiltration flow-through; and 5 is purified antibody.

[0020] Figure 4 It is the result of ELISA detection of multiple proteins based on the monoclonal antibody 1G6-60B12.

[0021] Figure 5These are the results of immunospot detection of multiple proteins based on the monoclonal antibody 1G6-60B12; 1 is BSA-p-Tau217; 2 is BSA-TAU; 3 is BSA; 4 is KLH-p-Tau217; 5 is TAU; 6 is KLH-Aβ; 7 is FT; and 8 is GFAP.

[0022] Figure 6 This is a graph showing the binding activity of monoclonal antibody 1G6-60B12 detected by ELISA. DETAILED DESCRIPTION

[0023] In order to enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments and drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts should fall within the scope of protection of the present invention.

[0024] Example 1: Acquisition of antigens

[0025] Sites 205 to 224 of the Tau protein were truncated and phosphorylated at site 217 to serve as the antigenic region. GenScript then synthesized the peptide and coupled the carrier protein KLH to the C-terminus of the short peptide for animal immunization and subsequent testing.

[0026] Example 2: Preparation of monoclonal antibodies

[0027] 1. Immunization of mice with p-Tau217 antigen

[0028] Step 1: After one week of acclimation, a 5- to 8-week-old female Balb / c mouse is rapidly mixed with an immune adjuvant and 50 μg of antigen, which is then injected intramuscularly into the hind leg to complete the first immunization. Three weeks later, the second immunization is performed in the same manner as the first immunization. Two weeks later, 50 μg of p-Tau217 antigen is diluted with PBS and injected intraperitoneally to complete the third immunization.

[0029] Step 2: ELISA to detect the titer of immunized mouse tail tip blood

[0030] Three days after the third immunization, tail blood was collected from the mice and centrifuged to collect serum, and its titer was detected by ELISA. The specific steps are as follows: add 0.1 μg / well of p-Tau217 antigen to the ELISA plate and coat for 2 hours; wash with TBST three times, 2 minutes each time; block with 200 μL / well of 3% BSA-TBST at 37°C for 2 hours; wash with TBST three times, 2 minutes each time; add diluted mouse serum (dilution factors are: 1000, 2000, 4000, 8000, 16000, 32000, 64000), 100 μL / well, incubate at 37°C for 1 hour; wash with TBST three times, 2 minutes each time; add goat anti-mouse IgG-HRP, 100 μL / well, incubate at 37°C for 45 minutes; wash with TBST three times, 2 minutes each time; add color development solution, 100 μL / well, incubate at room temperature for 10 minutes, stop with 2 mol / L sulfuric acid, and measure the absorbance on a microplate reader, where the measurement wavelength is 450 nm and the reference wavelength is 630 nm. Figure 1 The results of antibody titer testing of immunized mice, used for subsequent experiments, show that compared with unimmunized normal mice, the antibody titer of immunized mice was higher than 1:128,000. This result indicates that the immunized mice have produced an immune response to the immunogen p-Tau217 antigen and can be used for subsequent experiments.

[0031] 2. Cell fusion and screening of positive hybridoma cell lines

[0032] Step 1: Cell fusion

[0033] Myeloma SP2 / 0 cells were cultured in 1640 medium supplemented with 20% fetal bovine serum to a density of 1.0 × 10 7 ~1.0×10 8 Three days before fusion, 50 μg of p-Tau217 antigen was mixed with PBS and injected intraperitoneally to complete the pulse immunization. On the day of fusion, mice were sacrificed by cervical dislocation, and spleens were aseptically removed, triturated, and washed twice with serum-free medium. Splenocytes were mixed with SP2 / 0 cells at a ratio of 10:1 and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded, and the bottom of the centrifuge tube was gently tapped to loosen the cell pellet. Over 30 seconds, 1 mL of 45% PEG solution preheated to 37°C was slowly added. After standing at room temperature for 90 seconds, incomplete culture medium preheated to 37°C was slowly added. After mixing, the mixture was centrifuged at 800 rpm for 6 minutes. The supernatant was discarded, and 40 mL of complete culture medium preheated to 37°C was added. After mixing, the mixture was plated into a 96-well cell culture plate seeded with peritoneal macrophages and cultured in a 37°C, 5% CO2 incubator. From day 2 to 7 after fusion, HAT and HT were added every other day to select for fused cells.

[0034] Step 2: ELISA detection of positive hybridoma cells and cloning of hybridoma cells

[0035] Observe hybridoma cell growth 7-10 days after fusion. Hybridoma antibody secretion was assayed when cells in the wells exceeded one-third of the well bottom. The assay steps were as follows: add 0.1 μg / well of p-Tau217 antigen to the ELISA plate and coat for 2 hours; wash three times with TBST (2 minutes each time) and pat dry; block with 3% BSA-TBST at 37°C for 2 hours; wash three times with TBST (2 minutes each time) and pat dry; add 100 μL / well of hybridoma cell supernatant and incubate at 37°C for 1 hour; wash three times with TBST (2 minutes each time) and pat dry; add 100 μL / well of goat anti-mouse IgG-HRP and incubate at 37°C for 45 minutes; wash three times with TBST (2 minutes each time) and pat dry; add 100 μL / well of colorimetric solution and incubate at room temperature for 10 minutes. Incubate with 2 mol / L sulfuric acid and measure absorbance on a microplate reader at a wavelength of 450 nm and a reference wavelength of 630 nm. The clones with high OD values ​​were subcloned and screened again by ELISA 7-10 days later. After three rounds of cloning, one positive clone was finally screened and named 1G6-60B12.

[0036] 3. Identification of subtypes of monoclonal antibody 1G6-60B12

[0037] 0.1 μg / well of p-Tau217 antigen was added to the ELISA plate and coated for 2 hours; washed 3 times with TBST for 2 minutes each time; blocked with 3% BSA-TBST at 37°C for 2 hours; washed 3 times with TBST for 2 minutes each time; added hybridoma cell supernatant, 100 μL / well, incubated at 37°C for 1 hour; washed 3 times with TBST for 2 minutes each time; added HRP-labeled rabbit anti-mouse secondary antibody (IgG1, IgG2a, IgG2b, IgG2c, IgG3, IgM, kappa), 100 μL / well, incubated at 37°C for 45 minutes; washed 3 times with TBST for 2 minutes each time; added color development solution, 100 μL / well, incubated at room temperature for 10 minutes, terminated with 2 mol / L sulfuric acid, and measured absorbance on a microplate reader, where the measurement wavelength was 450 nm and the reference wavelength was 630 nm. The experimental results are as follows: Figure 2 As shown in the figure, the OD values ​​of anti-p-Tau217 IgG1 and kappa subtypes are much higher than those of other subtypes. Therefore, the heavy chain of monoclonal antibody 1G6-60B12 is IgG1 subtype, and the light chain is kappa subtype.

[0038] 4. Ascites Preparation and Antibody Purification

[0039] Step 1: Ascites preparation

[0040] Inject 500 μL of paraffin oil into the peritoneal cavity of 10-12 week-old Balb / c mice. After 7-10 days, culture the hybridoma cells to the logarithmic growth phase and adjust the cell density to 1.5×10 6~2×10 6 500 μL was injected intraperitoneally and ascites was collected 7 to 10 days later.

[0041] Step 2: Purification of monoclonal antibody 1G6-60B12

[0042] Binding: Filter the ascites and dilute to 40 mL with binding buffer (pH 7.0), add protein A beads, mix well, and bind on a shaker overnight at 4°C. Equilibration: Rinse the chromatography column three times with binding / wash buffer. Loading: Add the binding solution that has been bound overnight to the chromatography column, collect the flow-through and sample. Washing: After the protein A beads in the binding solution have settled to the bottom of the chromatography column, wash the chromatography column twice with binding / wash buffer. Elution: Elute the antibody bound to protein A with elution buffer (pH 3.0), sample, and immediately add 1 / 10 volume of neutralization buffer (pH 8.5) to the eluate and mix well. Ultrafiltration tube replacement: Concentrate the obtained elution neutralization solution using an ultrafiltration tube, and gradually replace the antibody into PBS (pH 7.2) using an ultrafiltration tube, and sample the first flow-through. After the concentration was determined by BCA method, the original ascites and samples from each step were subjected to SDS-PAGE electrophoresis and Coomassie brilliant blue staining. Figure 3 As shown in the figure, the heavy chain and light chain bands of the antibody can be clearly seen, and the impurity proteins are significantly reduced after purification, and the purity of the antibody is significantly improved.

[0043] Example 3: Application experiment of monoclonal antibodies

[0044] 1. ELISA detection of multiple proteins based on monoclonal antibody anti-1G6-60B12

[0045] The BSA-TAU, BSA-p-Tau217, KLH-p-Tau217, BSA, GFAP and FT proteins available in the laboratory were diluted to 1 μg / mL with coating buffer and added to the ELISA plate. The diluent was also added to the ELISA plate at 100 μL / well and coated for 2 h. The plate was washed 3 times with TBST for 2 min each time. 3% BSA-TBST, 200 μL / well, blocked at 37℃ for 2h; washed 3 times with TBST, 2 min / time; added the supernatant of the selected hybridoma cells 1G6-60B12, 100 μL / well, incubated at 37℃ for 1h; washed 3 times with TBST, 2 min / time; added goat anti-mouse IgG-HRP, 100 μL / well, incubated at 37℃ for 45min; washed 3 times with TBST, 2 min / time; added colorimetric solution, 100 μL / well, incubated at room temperature for 10min, terminated with 2mol / L sulfuric acid, and measured the absorbance on a microplate reader, where the measurement wavelength was 450nm and the reference wavelength was 630nm. Table 1 and Figure 4 The experimental results show that the monoclonal antibody 1G6-60B12 can specifically recognize the target antigen p-Tau217 short peptide.

[0046] Table 1 ELISA test results

[0047] Coating antigen BSA-TAU BSA-p-Tau217 KLH-p-Tau217 BSA GFAP FT OD 0.0146 1.5483 1.4245 0.0168 0.015 0.0157

[0048] 2. Immuno-spot detection of multiple proteins based on monoclonal antibody anti-1G6-60B12

[0049] Antigen coating: 0.5 μg of antigen BSA-p-Tau217, BSA-TAU, BSA, KLH-p-Tau217, TAU, KLH-Aβ, FT, and GFAP were spotted on the test paper with a distance between them; Primary antibody adsorption: 1 adsorption pad was added to the diluted primary antibody and incubated for 10 minutes; Sample incubation: The adsorbed primary antibody was added to the reaction well and incubated on a shaker at room temperature for 30 minutes at a frequency of 30 times / min; Washing: Washed 3 times with working solution for 3 minutes / time; Secondary antibody incubation: discard the working solution, add AP-labeled goat anti-mouse antibody reagent, 0.25mL / well, incubate on a shaker at room temperature for 30min, frequency 30 times / min; Washing: wash 3 times with working solution, 3min / time; Color development: aspirate the washing solution, add diluted reaction substrate, 0.25mL / well, and let it stand at room temperature for 4min; Stop color development: aspirate the reaction substrate, wash the test strip twice with distilled water to stop color development; Result analysis: aspirate the liquid in the reaction well and dry the test strip. Figure 5 The experimental results show that the monoclonal antibody anti-1G6-60B12 can specifically recognize the target antigen BSA-p-Tau217 and can be used to detect the p-Tau217 antigen in samples by immunospot method.

[0050] 3. ELISA to detect the binding activity of monoclonal antibodies against p-Tau217

[0051] ELISA plates were coated with BSA-p-Tau217 antigen at 0.1 μg / well and coated at 37°C for 2 h; washed with TBST three times for 2 min each; blocked with 3% BSA-TBST at 37°C for 2 h; washed with TBST three times for 2 min each; added with serially diluted monoclonal antibody anti-1G6-60B12 and incubated at 37°C for 1 h; washed with TBST three times for 2 min each; added with goat anti-mouse IgG-HRP at 100 μL / well and incubated at 37°C for 45 min; washed with TBST three times for 2 min each; added with colorimetric solution at 100 μL / well and incubated at room temperature for 15 min. The plates were then terminated with 2 mol / L sulfuric acid and the absorbance was measured on a microplate reader at a wavelength of 450 nm and a reference wavelength of 630 nm. Figure 6 The experimental results showed that the monoclonal antibody 1G6-60B12 can bind to the BSA-p-Tau217 antigen very well.

[0052] Example 4: Cloning and recombinant expression of monoclonal antibody variable region sequences

[0053] 1. Sequence determination of monoclonal antibody 1G6-60B12

[0054] Total RNA was extracted from hybridoma cells and reverse transcribed to obtain cDNA. The variable region fragments of the antibody heavy and light chains were amplified using the 5' RACE method. The amplified fragments were subcloned into the pEASY-Blunt vector, and the plasmids were extracted and sequenced. The sequence results of the light and heavy chains of the monoclonal antibody 1G6-60B12 were sequenced, and the complementarity determining regions of the antibody amino acid sequence were marked using the Kabat method. The results are shown in Table 2.

[0055] The amino acid sequence of the light chain variable region VL of the monoclonal antibody 1G6-60B12 is shown in SEQ ID No. 8, wherein the amino acid sequences of the three complementarity determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region are shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3, respectively, and the amino acid sequences of the framework regions LFR1, LFR2, LFR3 and LFR4 of the light chain variable region are shown in SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6 and SEQ ID No. 7, respectively.

[0056] The amino acid sequence of the heavy chain variable region VH of the monoclonal antibody 1G6-60B12 is shown in SEQ ID No. 16, wherein the amino acid sequences of the three complementarity determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region are shown in SEQ ID No. 9, SEQ ID No. 10 and SEQ ID No. 11, respectively, and the amino acid sequences of the framework regions HFR1, HFR2, HFR3 and HFR4 of the heavy chain variable region are shown in SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14 and SEQ ID No. 15, respectively.

[0057] Table 2

[0058]

[0059]

[0060] 2. Construction of recombinant plasmid of monoclonal antibody 1G6-60B12

[0061] The amplified fragment containing the 60B12-VH sequence was ligated with the linearized pCDH-mG1 vector via homologous recombination. The ligation product was transformed into E. coli DH5α competent cells, and the plasmid was extracted. Successful recombination was confirmed by sequencing and named pCDH-60B12-VH-mG1. The pCDH-60B12-VL-mk vector was constructed using the same steps as above.

[0062] 3. Recombinant expression of monoclonal antibody 1G6-60B12

[0063] The constructed pCDH-60B12-VH-mG1 and pCDH-60B12-VL-mk expression vectors were co-transfected into HEK293 cells for expression. Antibodies recognizing HpFT protein were obtained in the culture supernatant 72 hours after transfection. The target antibodies were purified from the culture supernatant using Protein A resin gel, and the activity of the recombinant antibodies was further tested by ELISA.

[0064] In summary, the present invention used BSA-p-Tau217 and BSA-TAU217 to screen monoclonal antibodies, and verified the specificity of the screened antibodies using multiple antigens. The results showed that the monoclonal antibody 1G6-60B12 of the present invention has good binding activity with p-Tau217 and strong specificity, and can be used for the detection of p-Tau217.

[0065] The above content is only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. An anti-p-Tau217 monoclonal antibody 1G6-60B12, characterized in that: The monoclonal antibody comprises a light chain and a heavy chain, the light chain is of kappa type and the heavy chain is of IgG1 type; wherein: The amino acid sequences of the three complementarity determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region are shown in SEQ ID No. 1, SEQ ID No. 2, and SEQ ID No. 3, respectively; The amino acid sequences of the three complementarity determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region are shown in SEQ ID No. 9, SEQ ID No. 10 and SEQ ID No. 11, respectively.

2. The anti-p-Tau217 monoclonal antibody 1G6-60B12 according to claim 1, characterized in that: The framework regions LFR1, LFR2, LFR3 and LFR4 of the light chain variable region have homologous sequences with at least 90% sequence identity to the amino acid sequences shown in SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6 and SEQ ID No. 7, respectively; The framework regions HFR1, HFR2, HFR3 and HFR4 of the heavy chain variable region have homologous sequences with at least 90% sequence identity to the amino acid sequences shown in SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14 and SEQ ID No. 15, respectively.

3. The anti-p-Tau217 monoclonal antibody 1G6-60B12 according to claim 1 or 2, characterized in that: The light chain variable region has a homologous sequence with at least 90% sequence identity to the amino acid sequence shown in SEQ ID No. 8; the heavy chain variable region has a homologous sequence with at least 90% sequence identity to the amino acid sequence shown in SEQ ID No.

16.

4. Use of the anti-p-Tau217 monoclonal antibody 1G6-60B12 according to any one of claims 1 to 3 in the preparation of a product for detecting the level of p-Tau217 antigen or a product for diagnosing a disease associated with the p-Tau217 antigen, wherein the disease is Alzheimer's disease.

5. A nucleic acid molecule, characterized in that: The nucleic acid molecule encodes the anti-p-Tau217 monoclonal antibody 1G6-60B12 according to any one of claims 1 to 3 or a functional fragment thereof.

6. A recombinant expression vector, characterized in that: The recombinant expression vector contains the nucleic acid molecule according to claim 5.

7. A host cell, characterized in that: The host cell contains the recombinant expression vector according to claim 6.

8. A product for detecting p-Tau217 antigen levels, characterized by: Comprising the nucleic acid molecule according to claim 5, the recombinant expression vector according to claim 6, and the host cell according to claim 7.

9. The product for detecting the level of p-Tau217 antigen according to claim 8, characterized in that: The recombinant expression vector has a signal peptide connected to the antibody and contains a transcriptional regulatory element; the host cell is selected from 293T cells, CHO cells, Expi293F TM Any type of cell.

10. The product for detecting the level of p-Tau217 antigen according to claim 8 or 9, characterized in that: The product includes any one of a detection reagent, a detection kit, and a detection test paper.

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