CN116375856A - anti-Tau protein monoclonal antibody 1A5-47H7, product based on same and application - Google Patents
anti-Tau protein monoclonal antibody 1A5-47H7, product based on same and application Download PDFInfo
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Abstract
The invention discloses an anti-Tau protein monoclonal antibody 1A5-47H7, a product based on the same and application thereof, and belongs to the technical field of monoclonal antibodies. The invention discloses the amino acid sequences of the complementarity determining regions of the variable regions of the light chain and the heavy chain of a monoclonal antibody against Tau protein, and experiments prove that the monoclonal antibody 1A5-47H7 against Tau protein has higher affinity and can specifically recognize target antigen Tau protein.
Description
Technical Field
The invention belongs to the technical field of monoclonal antibodies, and particularly relates to an anti-Tau protein monoclonal antibody (named 1A5-47H 7), a product based on the same and application thereof.
Background
Neurodegenerative diseases (neurogene-rate diseases) are a general term for diseases caused by degeneration and loss of chronic progressive cell neurons of brain and spinal cord, and Alzheimer's disease (alzheimer disease, AD) is one of the diseases related to age, which has the disease hidden, and the course of the disease is chronic, and is becoming a serious global health problem. Neurofibrillary tangles caused by hyperphosphorylation of microtubule-associated protein Tau (microtubule-associated protein Tau) are one of the main pathological features of AD, and are closely related to diagnosis of AD. The research shows that compared with normal people, the levels of Tau protein and P-Tau protein in cerebrospinal fluid and blood of AD patients are obviously increased, and the Tau protein can be used as an important biomarker of AD, and plays an important role in early discovery, early treatment and improvement of prognosis of AD.
Although there are many reports of anti-Tau antibodies at present, the reagents/kits based on the same for detecting Tau protein have the problems of insufficient sensitivity or low specificity. Therefore, a highly sensitive and highly specific method for detecting Tau protein content needs to be developed. The anti-Tau antibody with high specificity and high affinity is key to developing Tau protein immune detection reagents and kits.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide an anti-Tau protein monoclonal antibody, a product based on the anti-Tau protein monoclonal antibody and application thereof, so as to solve the defect that the reagent/kit for detecting the Tau protein in the prior art has insufficient sensitivity or low specificity.
In order to achieve the above purpose, the invention is realized by adopting the following technical scheme:
in a first aspect the invention provides an anti-Tau protein monoclonal antibody 1A5-47H7, the monoclonal antibody 1A5-47H7 comprising a light chain and a heavy chain, the light chain being of kappa and the heavy chain being of IgG2b; the amino acid sequences of 3 complementarity determining regions (LCDRs 1-3) of the light chain variable region of the monoclonal antibody 1A5-47H7 are respectively shown in SEQ ID No. 1-3, and the amino acid sequences of HCDRs 1-3 of the heavy chain variable region are respectively shown in SEQ ID No. 10-12.
Further, the monoclonal antibody 1A5-47H7 light chain variable region framework regions LFR 1-4 have at least 90% sequence identity with the amino acid sequences shown in SEQ ID Nos. 4-7, and the heavy chain variable region framework regions HFR 1-4 have at least 90% sequence identity with the amino acid sequences shown in SEQ ID Nos. 13-16.
Further, the monoclonal antibody 1A5-47H7 light chain variable region has a VL which has at least 90% sequence identity, preferably 95% sequence identity, to the amino acid sequence shown in SEQ ID No.8, and the heavy chain variable region has a VH which has at least 90% sequence identity, preferably 95% sequence identity, to the amino acid sequence shown in SEQ ID No. 17.
Further, the monoclonal antibody 1A5-47H7 light chain variable region has a VL that is at least 90% sequence identical, preferably 95% sequence identical, to the nucleotide sequence set forth in SEQ ID No.9, and the heavy chain variable region has a VH that is at least 90% sequence identical, preferably 95% sequence identical, to the nucleotide sequence set forth in SEQ ID No. 18.
In a second aspect, the invention provides a product for detecting Tau protein levels, comprising a monoclonal antibody or functional fragment thereof according to the first aspect of the invention, and any one of the following products:
a) Nucleic acid molecules: the nucleic acid molecule encodes a monoclonal antibody or a functional fragment thereof according to the first aspect of the invention;
b) Recombinant expression vector: the recombinant expression vector comprises the nucleic acid molecule described in a);
c) Host cell: said host cell comprising the recombinant expression vector described in b);
further, the recombinant expression vector has a signal peptide operably linked to an antibody;
further, the recombinant expression vector further comprises a transcriptional regulatory element.
Further, the product also includes a reagent that performs an antigen-antibody reaction or a reagent that performs a detection reaction;
still further, reagents for performing antigen-antibody reactions include buffers, salts, diluents, and the like;
a third aspect of the invention provides a method as claimed in any one of the following:
1) A method of making a monoclonal antibody according to the first aspect of the invention, the method comprising: culturing the host cell of the second aspect of the invention, optionally isolating the monoclonal antibody from the host cell and/or from the medium in which the host cell is grown;
2) A method for detecting Tau protein in a sample, said method comprising contacting a test sample with a monoclonal antibody according to the first aspect of the invention, and determining the level of Tau protein in said test sample.
Further, the method of 1) further comprises purifying the monoclonal antibody;
further, the host cell is selected from mammalian cells;
further, the cell is selected from 293T cells or CHO cells or Expi293FTM cells;
a fourth aspect of the invention provides the use of any one of the following:
1) The monoclonal antibody of the first aspect of the invention and the substance of the second aspect of the invention are applied to the detection of the Tau content;
2) The monoclonal antibody of the first aspect of the invention and the substance of the second aspect of the invention are applied to the preparation of products for detecting the Tau content;
3) The monoclonal antibody of the first aspect of the invention and the substance of the second aspect of the invention are used for preparing products for diagnosing Tau-related diseases;
further, the product includes a kit.
Further, the kit comprises: colloidal gold immunoassay kit, chemiluminescent detection kit, radioimmunoassay kit, enzyme-linked immunoassay kit, fluorescent immunoassay kit and microfluidic chip.
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses an anti-Tau protein monoclonal antibody (1A 5-47H 7), and specifically discloses the amino acid sequences of the complementarity determining regions of the variable regions of the light chain and the heavy chain of the monoclonal antibody.
Drawings
FIG. 1 shows results of a tail blood titer determination of Tau protein immunized mice;
FIG. 2 shows the results of the electrophoresis of monoclonal antibodies 1A5-47H 7; m, marker;1, ascites; 2, purifying;
FIG. 3 is a graph showing the results of the identification of the subtype 1A5-47H7 of monoclonal antibody;
FIG. 4 is the results of ELISA for the validation of monoclonal antibody 1A5-47H7 specificity;
FIG. 5 shows the result of western blot performed with monoclonal antibodies 1A5-47H 7; m, marker;1 and 3, U251 cells; 2 and 4, shsy5y cells;
FIG. 6 is the result of immunofluorescence detection of monoclonal antibodies 1A5-47H 7;
FIG. 7 is a graph showing the binding activity of monoclonal antibodies 1A5-47H7 detected by ELISA.
Detailed Description
In order that those skilled in the art will better understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
It should be noted that the terms "comprises" and "comprising," along with any variations thereof, in the description and claims of the present invention are intended to cover a non-exclusive inclusion, such as a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the attached drawing figures:
1. tau protein preparation
Synthesizing Tau gene sequence onto PCMV vector, and cloning target gene into PET-30a vector. Extracting plasmid, double enzyme cutting and identifying, and sequencing and comparing results. Transferring the constructed prokaryotic expression plasmid into competent cells of escherichia coli BL21 (DE 3), picking up monoclonal and inoculating the monoclonal into LB culture medium, shaking the bacteria at 37 ℃ until the OD value is 0.5-1, and adding IPTG and then carrying out overnight induction expression at 16 ℃. And (5) collecting thalli, and collecting a supernatant after ultrasonic crushing and centrifugation. And (3) dialyzing and concentrating the protein purified by Ni column affinity chromatography to obtain the high-concentration Tau protein.
2. Acquisition of hybridoma cells
Step one, tau protein immunization of mice
3 female Balb/c mice of 5-8 weeks of age; after one week of adaptation feeding, 50 mug of Tau protein is mixed with Freund's complete adjuvant 1:1, and the mixture is completely emulsified to a water-in-oil state and then injected subcutaneously at 4 points to complete the first immunization; after three weeks, 50 mug of Tau protein is mixed with Freund's incomplete adjuvant 1:1, and the mixture is completely emulsified to a water-in-oil state and then injected subcutaneously for 4 points to complete the second immunization; three weeks later, 50 μg of Tau protein was diluted with PBS and injected intraperitoneally to complete the third immunization.
Step two, ELISA detection of blood titer of the tip of the tail of the immune mouse
Three weeks after the third immunization, blood serum was collected from the rat tail and assayed for potency by ELISA. The method comprises the following specific steps: after the Tau protein is diluted to a proper concentration, the Tau protein is added into an ELISA plate, 100 ng/hole is added, and the ELISA plate is coated overnight at 4 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; 3% BSA-PBST 200 ul/well blocked at 37℃for 2h; PBST is washed for 3 times, 2 min/time and is patted dry; adding mouse serum diluted in a gradient manner according to a certain proportion, and incubating for 1h at 37 ℃ at 100 mu L/hole; PBST is washed for 3 times, 2 min/time and is patted dry; goat anti-mouse IgG-HRP was added, 100. Mu.L/well and incubated at 37℃for 40min; PBST is washed for 3 times, 2 min/time and is patted dry; after adding the color development solution, 100. Mu.L/well and incubating at Room Temperature (RT) for 10min, the stop solution was stopped and the absorbance was measured on an ELISA, wherein the measurement wavelength was 450nm and the reference wavelength was 630nm. Figure 1 gives the results of the immune mouse antibody titer test for the subsequent experiments, which shows that: compared with normal mice which are not immunized, the titer of the immunized mice is higher than 1:12.8 ten thousand. This result demonstrates that the immunized mice developed an immune response to the immunogen Tau protein and can be used in subsequent experiments.
Step three, cell fusion
Myeloma cells SP2/0 were adapted to be fed with 20% fetal bovine serum for one week. Taking 1 immunized mouse, taking 50 mug Tau protein after fusion for 3 days, evenly mixing with PBS, and performing intraperitoneal injection to complete impact immunization. On the day of fusion, mice were sacrificed aseptically to harvest the spleens, and after milling, washed 2 times with serum-free medium. Spleen cells were mixed with SP2/0 cells at a ratio of 10:1, centrifuged at 1200rpm for 6min and the supernatant was discarded. Flick the bottom of the centrifuge tube to loosen the cell pellet. 1mL of 45% PEG solution preheated to 37℃was slowly added over 30 s. After standing at room temperature for 90s, the incomplete medium preheated to 37℃was slowly added. After mixing, the mixture was centrifuged at 800rpm for 6min, and the supernatant was discarded. Adding 40mL of complete culture medium preheated to 37deg.C, mixing, adding into 96-well cell culture plate inoculated with abdominal cavity macrophage, placing at 37deg.C, 5% CO 2 Culturing in an incubator. The selection of fused cells was performed by supplementing HAT and HT on days 2-7 after fusion.
Step four, ELISA detection of positive hybridoma cells and cloning of hybridoma cells
And (3) after 7-10 days of fusion, observing the growth condition of the hybridoma cells, and detecting the secretion condition of the hybridoma antibodies when the cells in the holes exceed 1/3 of the bottoms of the holes. The detection steps are as follows: after the Tau protein is diluted to a certain concentration, the Tau protein is added into an ELISA plate, 100 mu L/hole is added, and the ELISA plate is coated overnight at 4 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; blocking for 2h at 37℃with 3% BSA-PBST; PBST is washed for 3 times, 2 min/time and is patted dry; adding hybridoma cell supernatant, 100 mu L/well, and 37 ℃ for 1h; PBST is washed for 3 times, 2 min/time and is patted dry; goat anti-mouse IgG-HRP (goat anti-mouse IgG-HRP) was added at 100. Mu.L/well for 40min at 37 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; after adding the color development solution, 100. Mu.L/well and incubating for 10min at RT (room temperature), the stop solution was stopped and the absorbance was measured on a microplate reader, wherein the measurement wavelength was 450nm and the reference wavelength was 630nm. Clones with high OD values were selected for subcloning and ELISA screening was performed again after 7-10 days. After 3 clones, 1 positive clone was finally selected and designated 1A5-47H7.
3. Preparation of monoclonal antibody 1A5-47H7 from ascites
mu.L of ascites adjuvant was injected into the peritoneal cavity of 10 week old Balb/c mice. After 7 days, the hybridoma cells were culturedTo logarithmic phase, cell density was adjusted to 2X 10 6 mu.L was injected intraperitoneally at a volume of 500. Mu.L. Ascites was collected after 7-10 days.
4. Purification of monoclonal antibody 1A5-47H7
Combining: the ascites was diluted with binding/wash buffer (pH 7.0), centrifuged to collect the supernatant, and protein A beads were added and mixed well and allowed to bind overnight on a shaker at 4 ℃. Balance: binding/wash buffer equilibrates the column 2 times. And (3) column loading: the overnight bound conjugate was applied to a chromatographic column. Washing: after protein A beads in the binding solution are sunk at the bottom of the chromatographic column, the chromatographic column is washed for 2 times by using binding/wash buffer. Eluting: the antibody bound to protein A was eluted with an Elutation buffer (pH 3.0), and 1/10 of the volume of the neutralization buffer (pH 8.5) was immediately added to the eluate and mixed. Ultrafiltration displacement: the elution neutralization solution obtained in the previous step was concentrated by ultrafiltration tube, and the solution was gradually replaced into antibody storage solution PBS (pH 7.2). After the concentration is measured by BCA method, a proper amount of SDS-PAGE electrophoresis and Coomassie blue staining are carried out simultaneously with the original ascites, the result is shown in figure 2, the heavy chain and light chain bands of the antibody can be clearly seen, the impurity protein is obviously reduced after purification, and the purity of the antibody is obviously improved.
5. Identification of monoclonal antibody 1A5-47H7 subtype
100 ng/hole of Tau protein is added to the ELISA plate, and the ELISA plate is coated overnight at 4 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; blocking for 2h at 37℃with 3% BSA-PBST; PBST is washed for 3 times, 2 min/time and is patted dry; adding hybridoma cell supernatant, 100 mu L/hole, and incubating at 37 ℃ for 1h; PBST is washed for 3 times, 2 min/time and is patted dry; HRP-labeled rabbit anti-mouse secondary antibodies (IgG 1, igG2a, igG2b, igG3, igM, kappa) were added, 100. Mu.L/well and incubated at 37℃for 40min; PBST is washed for 3 times, 2 min/time and is patted dry; after adding the chromogenic solution and incubating for 10min at 100. Mu.L/well at RT, the stop solution was stopped and the absorbance was measured on an ELISA reader, wherein the measurement wavelength was 450nm and the reference wavelength was 630nm. As shown in FIG. 3, the OD values of the 1A5-47H7 IgG2b and kappa type were much higher than those of the other subtypes and negative control. Thus, the 1A5-47H7 monoclonal antibody is of the IgG2b type in heavy chain and of the kappa type in light chain.
6. ELISA for verifying specificity of monoclonal antibody 1A5-47H7
The purified Tau protein expressed in example 1 and the laboratory-existing NFL and BSA proteins were diluted to 1 μg/mL with coating buffer, respectively, added to the ELISA plate, and the dilutions were added to the ELISA plate as well, 100 μl/well, overnight at 4 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; 3% BSA-PBST 200 ul/well blocked at 37℃for 2h; PBST is washed for 3 times, 2 min/time and is patted dry; adding the supernatant of hybridoma cell 1A5-47H7, 100 mu L/hole, and incubating at 37 ℃ for 1H; PBST is washed for 3 times, 2 min/time and is patted dry; goat anti-mouse IgG-HRP was added, 100. Mu.L/well and incubated at 37℃for 40min; PBST is washed for 3 times, 2 min/time and is patted dry; after adding the chromogenic solution and incubating for 10min at 100. Mu.L/well at RT, the stop solution was stopped and the absorbance was measured on an ELISA reader, wherein the measurement wavelength was 450nm and the reference wavelength was 630nm.
Experimental results (table 1 and fig. 4) illustrate: monoclonal antibody 1A5-47H7 can specifically recognize target antigen Tau protein.
TABLE 1ELISA to verify the specificity of monoclonal antibodies 1A5-47H7
7. Application of monoclonal antibody 1A5-47H7 in western blot
After the total proteins of U251 and SH-SY5Y cells were collected and quantified by the BCA method, 20. Mu.g of the total proteins were subjected to SDS-PAGE, and after the completion, PVDF membrane was transferred. After blocking with 5% nonfat dry milk, the purified 1A5-47H7 was incubated. After 3 washes of TBST, sheep anti-mouse IgG-HRP was incubated. After 3 washes of TBST, ECL developed. The results (FIG. 5) show that in the 50-80 kDa band interval, tau-specific bands appear in the cellular protein samples, and that 1A5-47H7 is consistent with the commercial anti-Tau antibody bands. This result demonstrates that 1A5-47H7 is capable of specifically recognizing Tau protein and can be used for western blot to detect Tau protein in a sample.
8. Application of monoclonal antibody 1A5-47H7 in immunofluorescence detection
Inoculating U251 and SHSY5Y cells to the climbing slices, fixing with paraformaldehyde, washing with PBST 3 times, incubating with 3% BSA-TBST (containing 0.1% Triton 100) RT (room temperature) for 1h, washing with PBST once, and incubating with RT for 1A5-47H7,2h; after three PBST washes, the Alexa Fluor 549-labeled goat anti-mouse IgG antibody was incubated at RT for 60min. After PBST washing, DAPI-containing caplets were added. The staining results were observed under a fluorescence microscope. The experimental results (fig. 6) show that after 2 cells are incubated by the antibody, obvious red (fig. 6 Tau) and blue nuclei stained by DAPI (fig. 6 DAPI) can be observed under a microscope, and the two nuclei can be well overlapped (fig. 6 DAPI/Tau), and the results indicate that 1A5-47H7 can specifically stain Tau protein and can be used for immunofluorescent staining to detect Tau protein in a sample.
9. ELISA detection of monoclonal antibody 1A5-47H7 binding Activity
Tau protein coated ELISA plate, 100 ng/well, overnight at 4 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; blocking for 2h at 37℃with 3% BSA-PBST; PBST is washed for 3 times, 2 min/time and is patted dry; adding 1A5-47H7 after gradient dilution, and incubating for 1H at 37 ℃; PBST is washed for 3 times, 2 min/time and is patted dry; goat anti-mouse IgG-HRP was added, 100. Mu.L/well and incubated at 37℃for 40min; PBST is washed for 3 times, 2 min/time and is patted dry; after adding the chromogenic solution and incubating for 10min at 100. Mu.L/well at RT, the stop solution was stopped and the absorbance was measured on an ELISA reader, wherein the measurement wavelength was 450nm and the reference wavelength was 630nm. The experimental results (fig. 7) demonstrate that: monoclonal antibody 1A5-47H7 can well bind Tau protein and presents concentration dependence, EC 50 0.018 μg/mL.
10. Monoclonal antibody 1A5-47H7 sequencing
Extracting total RNA of the hybridoma cells, and carrying out reverse transcription to obtain cDNA; amplifying antibody heavy chain and light chain variable region fragments by a 5' RACE method; subcloning the amplified fragment into pEASY-Blunt vector, extracting plasmid and sequencing; the result of the sequenced antibody light and heavy chain sequences (SEQ ID Nos. 8-9 and 17-18) was followed by labelling the CDR regions of the amino acid sequences of the antibodies using the Kabat method.
1A5-47H7 antibody light chain variable region (VL) amino acid sequence (SEQ ID No. 8):
1A5-47H7 antibody light chain variable region (VL) gene sequence (SEQ ID No. 9):
1A5-47H7 antibody heavy chain variable region (VH) amino acid sequence (SEQ ID No. 17):
1A5-47H7 antibody heavy chain variable region (VH) gene sequence (SEQ ID No. 18):
in conclusion, the monoclonal antibody 1A5-47H7 provided by the invention has better binding activity with Tau, has strong specificity, and can be used for detecting Tau.
The above is only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited by this, and any modification made on the basis of the technical scheme according to the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Claims (10)
1. A monoclonal antibody against Tau protein comprising a light chain and a heavy chain, the light chain being kappa and the heavy chain being IgG2b; wherein:
the amino acid sequences of the 3 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 3 complementarity determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region are shown in SEQ ID No.10, SEQ ID No.11 and SEQ ID No.12, respectively.
2. The monoclonal antibody against Tau protein of claim 1, wherein the amino acid sequences of the variable light chain framework regions LFR1, LFR2, LFR3 and LFR4 of the monoclonal antibody against Tau protein are shown in SEQ ID No.4, SEQ ID No.5, SEQ ID No.6 and SEQ ID No.7, respectively;
the amino acid sequences of heavy chain variable region framework regions HFR1, HFR2, HFR3 and HFR4 of the anti-Tau protein monoclonal antibody are respectively shown as SEQ ID No.13, SEQ ID No.14, SEQ ID No.15 and SEQ ID No. 16.
3. The monoclonal antibody against Tau protein of claim 1 or 2, wherein the light chain variable region of the monoclonal antibody against Tau protein has at least 90% sequence identity with the amino acid sequence shown in SEQ ID No. 8; the heavy chain variable region of the monoclonal antibody against Tau protein has at least 90% sequence identity with the amino acid sequence shown in SEQ ID No. 17.
4. A monoclonal antibody against the Tau protein of claim 3, wherein the light chain variable region of the monoclonal antibody has 95% sequence identity to the amino acid sequence shown in SEQ ID No. 8; the heavy chain variable region of the monoclonal antibody against Tau protein has 95% sequence identity with the amino acid sequence shown in SEQ ID No. 17.
5. The monoclonal antibody against Tau protein of claim 1 or 2, wherein the light chain variable region of the monoclonal antibody against Tau protein has at least 90% sequence identity with the nucleotide sequence shown in SEQ ID No. 9; the heavy chain variable region of the anti-Tau protein monoclonal antibody has at least 90% sequence identity with the nucleotide sequence shown in SEQ ID No. 18.
6. The monoclonal antibody against Tau protein of claim 5, wherein the light chain variable region of the monoclonal antibody against Tau protein has 95% sequence identity with the nucleotide sequence shown in SEQ ID No. 9; the heavy chain variable region of the monoclonal antibody against Tau protein has 95% sequence identity with the nucleotide sequence shown in SEQ ID No. 18.
7. A product for detecting Tau protein levels comprising:
a) A nucleic acid molecule encoding the monoclonal antibody or functional fragment thereof according to any one of claims 1 to 6;
b) A recombinant expression vector comprising the nucleic acid molecule of a);
c) A host cell comprising the recombinant expression vector of b).
8. The use of the monoclonal antibody against Tau protein according to any one of claims 1 to 6 and the product for detecting Tau protein level according to claim 7 for the preparation of a product for detecting Tau protein, characterized in that the product comprises a detection reagent, a kit or a test strip.
9. Use of the monoclonal antibody against Tau protein according to any one of claims 1 to 6 and the product for detecting Tau protein level according to claim 7 for the preparation of a product for diagnosing a disease associated with Tau protein, characterized in that said product comprises a diagnostic reagent, a kit or a diagnostic test paper.
10. The use according to claim 9, wherein the Tau protein associated disease is a neurodegenerative disease.
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| CN116925217A (en) * | 2023-09-14 | 2023-10-24 | 北京凯祥弘康生物科技有限公司 | Antibodies to Tau protein |
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