Thyroid stimulating hormone antibody and application thereof in fluorescence immunochromatography detection

Abstract

The invention provides a thyroid stimulating hormone antibody and application thereof in fluorescence immunochromatography detection. A heavy chain CDR1 sequence of the monoclonal antibody 8I22 is shown as SEQ ID NO: 1, a heavy chain CDR2 sequence of the monoclonal antibody 8I22 is shown as SEQ ID NO: 2, and a heavy chain CDR3 sequence of the monoclonal antibody 8I22 is shown as SEQ ID NO: 3; the sequence of the light chain CDR1 is as shown in SEQ ID NO: 4, the sequence of the light chain CDR2 is as shown in SEQ ID NO: 5, and the sequence of the light chain CDR3 is as shown in SEQ ID NO: 6. A heavy chain CDR1 sequence of the monoclonal antibody 9E6 is shown as SEQ ID NO: 11, a heavy chain CDR2 sequence of the monoclonal antibody 9E6 is shown as SEQ ID NO: 12, and a heavy chain CDR3 sequence of the monoclonal antibody 9E6 is shown as SEQ ID NO: 13; the sequence of the light chain CDR1 is as shown in SEQ ID NO: 14, the sequence of the light chain CDR2 is as shown in SEQ ID NO: 15, and the sequence of the light chain CDR3 is as shown in SEQ ID NO: 16. The antibody is high in sensitivity, low in detection limit and wide in linear range when being used for detecting thyroid stimulating hormone.

Description

Technical Field

[0001] The present invention relates to a thyroid stimulating hormone antibody and application thereof in fluorescent immunochromatographic detection, belonging to the technical field of thyroid stimulating hormone detection. Background Art

[0002] Thyroid hormone (TSH) is a glycoprotein hormone secreted by the pituitary gland. It consists of two subunits, α and β, with the β subunit possessing specific biological activity. TSH regulates the synthesis and secretion of thyroid hormones (T3 / T4) by binding to TSH receptors on the membrane of thyroid follicular cells. It is a core regulator of the hypothalamic-pituitary-thyroid axis (HPT axis). Dynamic changes in TSH levels can sensitively reflect thyroid function. Its detection is of great clinical value in the diagnosis and treatment of hyperthyroidism, hypothyroidism, subclinical thyroid disease, and postoperative monitoring of thyroid cancer. According to the "Guidelines for the Diagnosis and Treatment of Thyroid Diseases in China," the normal reference range for serum TSH is 0.3-5.0 mIU / L. Abnormal serum TSH concentrations (e.g., elevated TSH indicates primary hypothyroidism, and decreased TSH indicates hyperthyroidism) are the preferred screening marker for thyroid dysfunction.

[0003] Serum TSH testing technology has undergone four generations of innovation: the first-generation radioimmunoassay (RIA) had low sensitivity (1-2 mIU / L), enabling only the diagnosis of hypothyroidism; the second-generation immunoradiometric assay (IRMA) improved sensitivity to 0.1-0.2 mIU / L, enabling the diagnosis of hyperthyroidism; the third-generation chemiluminescence assay (ICMA) and the fourth-generation time-resolved fluorescence assay (TRIFA) have advanced sensitivity to 0.001-0.002 mIU / L, enabling the identification of subclinical thyroid dysfunction. However, current mainstream testing methods (such as CLIA and ELISA) rely on large equipment and are complex to operate, making them difficult to meet the needs of primary care and point-of-care rapid testing (POCT). Fluorescence immunochromatography (FICA), based on the principle of specific antigen-antibody binding and incorporating signal amplification technologies such as quantum dots and rare earth fluorescent microspheres, offers advantages such as rapid testing (<15 minutes), small sample volume (10 μL of whole blood), and resistance to matrix interference. It is particularly suitable for accurate testing of complex samples (such as hemolysis and lipemia), becoming a key technology for promoting TSH diagnosis at the grassroots level. The development of a TSH detection kit based on fluorescent immunochromatography could overcome traditional technical bottlenecks and provide a highly efficient and portable detection tool for clinical use, with significant clinical application value and market potential. However, existing technologies currently lack TSH antibodies that can be used in fluorescent immunochromatography. Summary of the Invention

[0004] The present invention provides a thyroid stimulating hormone antibody and its application in fluorescent immunochromatographic detection, which can effectively solve the above problems.

[0005] A thyroid-stimulating hormone monoclonal antibody 8I22, The heavy chain CDR1 sequence of the monoclonal antibody 8I22 is shown in SEQID NO: 1, the heavy chain CDR2 sequence is shown in SEQID NO: 2, and the heavy chain CDR3 sequence is shown in SEQID NO: 3; the light chain CDR1 sequence is shown in SEQID NO: 4, the light chain CDR2 sequence is shown in SEQID NO: 5, and the light chain CDR3 sequence is shown in SEQID NO: 6.

[0006] In some embodiments, the heavy chain variable region sequence of the monoclonal antibody 8I22 is shown in SEQ ID NO: 7, and the light chain variable region sequence is shown in SEQ ID NO: 8.

[0007] In some embodiments, the heavy chain sequence of the monoclonal antibody 8I22 is shown in SEQ ID NO:9, and the light chain sequence is shown in SEQ ID NO:10.

[0008] A thyroid-stimulating hormone monoclonal antibody 9E6, wherein the heavy chain CDR1 sequence of the monoclonal antibody 9E6 is shown in SEQ ID NO: 11, the heavy chain CDR2 sequence is shown in SEQ ID NO: 12, and the heavy chain CDR3 sequence is shown in SEQ ID NO: 13; the light chain CDR1 sequence is shown in SEQ ID NO: 14, the light chain CDR2 sequence is shown in SEQ ID NO: 15, and the light chain CDR3 sequence is shown in SEQ ID NO: 16.

[0009] In some embodiments, the heavy chain variable region sequence of the monoclonal antibody 9E6 is shown in SEQ ID NO: 17, and the light chain variable region sequence is shown in SEQ ID NO: 18.

[0010] In some embodiments, the heavy chain sequence of the monoclonal antibody 9E6 is shown in SEQ ID NO: 19, and the light chain sequence is shown in SEQ ID NO: 20.

[0011] A fluorescent immunochromatographic detection agent for thyroid stimulating hormone, comprising: TSH fluorescent immunochromatographic detection microspheres and capture antibodies; The TSH fluorescent immunochromatographic detection microspheres are prepared by first preparing avidin-labeled fluorescent microspheres; labeling the monoclonal antibody 9E6 with biotin to obtain biotin-labeled monoclonal antibody 9E6; and then mixing the avidin-labeled fluorescent microspheres and the biotin-labeled monoclonal antibody 9E6 to obtain the TSH fluorescent immunochromatographic detection microspheres. The capture antibody is the monoclonal antibody 8I22.

[0012] A fluorescent immunochromatographic detection agent card for thyroid stimulating hormone comprises the fluorescent immunochromatographic detection agent for thyroid stimulating hormone.

[0013] In some embodiments, the fluorescent immunochromatographic detection card for thyroid stimulating hormone comprises absorbent paper, nitrocellulose membrane, conjugate pad and blank substrate; The TSH fluorescent immunochromatographic detection microspheres are provided on the conjugate pad; The nitrocellulose membrane is coated with a detection T line and a quality control C line, wherein the detection T line is a capture antibody and the quality control C line is an avidin polyclonal antibody.

[0014] In some embodiments, the coating amount of the detection T line is 0.5-1.0 mg / mL; the coating amount of the quality control C line is 0.2-0.5 mg / mL.

[0015] The beneficial effects of the present invention are: The present invention addresses the limited sensitivity of lateral flow chromatography technology compared to chemiluminescence. By optimizing highly sensitive antibody pairs and integrating a biotin-avidin signal amplification system, a fluorescence chromatography detection process with high detection sensitivity, low detection limit and wide linear range is developed. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0017] Figure 1 This is the consistency curve between the test value of the reagent card in the embodiment of the present invention and the Roche assignment value.

[0018] Figure 2 Schematic diagram of a reagent card according to an embodiment of the present invention. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are 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 work are within the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention.

[0020] Example 1 Preparation of monoclonal antibodies 1. Obtaining TSH monoclonal antibodies Mice were immunized with recombinant human thyroid-stimulating hormone (TSH), and the resulting hybridomas were screened to obtain hybridomas that secrete high-affinity antibodies to human thyroid-stimulating hormone (TSH). Antibodies that cross-react with human luteinizing hormone (LH), human follicle-stimulating hormone (FSH), and human chorionic gonadotropin (hCG) were excluded. Monoclonal antibodies were prepared using conventional methods.

[0021] 2. Obtaining the heavy and light chain gene sequences of anti-TSH monoclonal antibodies RNA was extracted from the selected hybridoma cells, and reverse transcription was performed using the RNA as a template to obtain cDNA, which was cloned into a vector and then sequenced; Analysis of the variable region genes of the light and heavy chains of the antibodies: Based on the sequencing results, the VH and VL as well as the constant regions were analyzed, and the sequences of antibodies 8I22 and 9E6 were finally obtained as follows: The sequence of antibody 8I22 is as follows: Heavy chain CDR1: GFSLSTY (SEQ ID NO: 1) Heavy chain CDR2: VIWAGGSTDY (SEQ ID NO: 2) Heavy chain CDR3: CAREGGYAMDY (SEQ ID NO: 3) Light chain CDR1: RASQDISNYL (SEQ ID NO: 4) Light chain CDR2: GASTLQS (SEQ ID NO: 5) Light chain CDR3: QQYNSYPYT (SEQ ID NO: 6) Heavy chain variable region: QVQLQQSGPGLVKPSQTLSLTCTVSGFSLSTYGVGWVRQPPGKGLEWLGVIWAGGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCAREGGYAMDYWGQGTLVTVSS (SEQ ID NO:7) Light chain variable region: DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKAPKLLIYGASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKVEIK (SEQ ID NO:8) Heavy chain: QVQLQQSGPGLVKPSQTLSLTCTVSGFSLSTYGVGWVRQPPGKGLEWLGVIWAGGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCAREGGYAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPVK (SEQ ID NO:9) Light chain: DIQMTQSPSSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKAPKLLIYGASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10) The sequence of antibody 9E6 is as follows: Heavy chain CDR1: GYSFTS (SEQ ID NO: 11) Heavy chain CDR2: IYPGGGDT (SEQ ID NO: 12) Heavy chain CDR3: CARDYGDY (SEQ ID NO: 13) Light chain CDR1: RASQSVGNNKLA (SEQ ID NO: 14) Light chain CDR2: GASSRAT (SEQ ID NO: 15) Light chain CDR3: QQYGSSPTT (SEQ ID NO: 16) Heavy chain variable region: QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYGISWVRQAPGQGLEWMGGIYPGGGDTKYNEKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARDYGDYWGQGTLVTVSS (SEQID NO: 17) Light chain variable region: EIVLTQSPGTLSLSPGERATLSCRASQSVGNNKLAWYQQKPGQAPRLLIYGASSRATGVPSRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYGSSPTTFGAGTKLELK (SEQ ID NO: 18) Heavy chain: QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYGISWVRQAPGQGLEWMGGIYPGGGDTKYNEKFKGKATLTADKSSSTAYMQLSSSLTSEDSAVYYCARDYGDYWGQGTLVTV SSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTK GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPVK (SEQID NO:19) Light chain: EIVLTQSPGTLSLSPGERATLSCRASQSVGNNKLAWYQQKPGQAPRLLIYGASSRATGVPSRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYGSSPTTFGAGTKLELKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20) Example 2: Preparation of TSH fluorescent immunochromatographic detection microspheres 1. Labeling of Avidin (SA) Fluorescent Microspheres 1) Microsphere Assay: Take 20 μL of microspheres (purchased from Weidu Biotechnology, Catalog No. FT0300CC, 10 mg / mL concentration). Label with a ratio of 0.2 mg microspheres to 0.02 mg SA (mass ratio 10:1).

[0022] 2) Microsphere activation: Add 200 μL of 0.05 M MES and 20 μL of 1 mg / mL EDC to the microspheres, sonicate for 30 seconds, and shake in a constant temperature shaker at room temperature for 5 minutes; 3) Coupling: Add 20 μg SA of the antibody to be labeled to the system, sonicate for 30 seconds, and shake in a constant temperature oscillator at room temperature for 15 minutes; 4) Blocking: Add 10 μL of 20% BSA to the system, sonicate for 30 seconds, and shake in a constant temperature shaker at room temperature for 10 minutes; 5) Centrifugation: After the reaction is complete, centrifuge at 15,000 rpm and 4°C for 15 minutes. 6) Resuspension: After centrifugation, carefully remove the supernatant and add 100 μL of microsphere resuspension buffer (50 mM Tris-HCl, 0.5% BSA, 0.2% Tween-20, 0.05% Proclin 300) to resuspend the pellet to 5 times the volume of the microspheres.

[0023] 2. Biotinylation of Detection Monoclonal Antibody 9E6 1) Calculate the amount of antibody and biotin required for labeling based on a molar ratio of Ab:Bio = 1:5.

[0024] 2) Add the calculated amount of biotin to the antibody, mix thoroughly, and incubate in a constant temperature shaker at room temperature for 2 hours.

[0025] 3) After the reaction is complete, add 10% glycine at a volume ratio of 50:1, mix thoroughly, and shake in a constant temperature shaker at room temperature for 30 minutes.

[0026] 4) After the reaction is completed, use dialysis to remove the biotin labeled in the system.

[0027] 3. Biotinylated 9E6-bio antibody binds to avidin microspheres 1) Take 200 μL of fluorescent microspheres pre-labeled with avidin, sonicate for 30 seconds, add 9E6-bio at a concentration ratio of 1:20, and shake in a constant temperature shaker at room temperature for 30 minutes; 2) After the reaction, centrifuge at 15,000 rpm and 4°C for 15 minutes; 3) After centrifugation, carefully remove the supernatant and add 100 μL of microsphere resuspension buffer (50 mM Tris-HCl, 0.5% BSA, 0.2% Tween-20, 0.05% Proclin 300) to resuspend the microspheres to the original volume. This is the TSH fluorescent immunochromatographic detection microsphere.

[0028] Example 3: Preparation of TSH fluorescent immunochromatographic reagent card 1. Preparation of fluorescent marker conjugate pad: The TSH fluorescent immunochromatographic detection microspheres (9E6-bio antibody labeled fluorescent microspheres) prepared in Example 2 were diluted 50 times with the microsphere resuspension solution and sprayed onto a 10 mm wide glass fiber mat at 60 μL / cm. The sprayed labeled mat was then placed in a 37°C oven to dry for more than 2 hours. After drying, the sprayed labeled mat was placed in an aluminum foil bag pre-added with desiccant and sealed for later use. The TSH fluorescent immunochromatographic detection microspheres were 2. Detect the coating of T line and quality control C line: Dilute the TSH capture antibody 8I22 to 0.5-1.0 mg / mL in 10 mM PBS buffer (pH 7.4) as the test T line; dilute the rabbit anti-SA polyclonal antibody to 0.4 mg / mL as the quality control C line. Coat the test T line and quality control C line onto the nitrocellulose membrane on the back plate at a coating volume of 1.0 μL / cm. Dry in a 37°C oven for 30 hours. After drying, seal the membrane in an aluminum foil bag pre-filled with desiccant and store until ready for use. 3. Assembly and cutting of test reagent cards At a humidity below 25%, take absorbent paper and a blank glass fiber pad and cut them into 30mm widths. Take out the treated coated plate and conjugation pad, and arrange the components from top to bottom in the order of absorbent paper-nitrocellulose membrane-conjugation pad-blank glass fiber. The absorbent paper and conjugation pad are pressed on the nitrocellulose membrane with a width of about 1-2mm, and then the blank glass fiber is pasted and pressed on the conjugation pad with a width of about 1-2mm. The assembled large plate is cut into 4mm widths as a TSH fluorescent immunochromatography reagent card. Figure 2 shown.

[0029] Example 4: 1. Reagent card linearity test Test method: Take Roche assigned clinical sample, mix 10 μL with 90 μL sample buffer (20 mM Tris-HCl, 0.3% CN, 0.2% Tween-20, 0.05% S9, 0.2% trehalose, 0.05% Proclin300), take 80 μL and add the sample. At the 15th minute of the reaction, use a fluorescent immunoassay to detect the fluorescence values of the T line and C line.

[0030] The test results are shown in Table 1. As shown in Table 1, the consistency between the back-calculated concentration and the Roche assigned concentration can reach 0.996, and the minimum detection limit is below 0.02mIU / L.

[0031] Table 1 TSH Roche assigned sample conformity test

[0032] 2. Reagent card linear precision (CV) test Test method: The test steps are the same as above. Use samples in the low-value range (0.02-0.14mIU / L) assigned by Roche, and test three replicates for each sample.

[0033] The test results are shown in Table 2. As shown in Table 2, the limit of quantification of this method can reach 0.02mIU / L.

[0034] Table 2 TSH Roche assigned sample conformity test

[0035] 3. Cross-reactivity test with LH, FSH and HCG Test Method: Dilute LH and FSH standards in assay buffer to a concentration of 200mIU / L, and HCG to 1000mIU / L. A pass criterion is a back-calculated concentration <0.01mIU / L.

[0036] The test steps were the same as above, and four parallel tests were performed for each antigen.

[0037] The results are shown in Table 3. As can be seen from Table 3, the calculated concentrations of the three antigens are all lower than 0.01 mIU / L, thus confirming that the reagent card does not have any cross-reaction with these three glycoprotein hormones.

[0038] Table 3 Cross-talk with LH, FSH and HCG

[0039] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A thyroid stimulating hormone monoclonal antibody 8I22, characterized in that The heavy chain CDR1 sequence of the monoclonal antibody 8I22 is shown in SEQID NO: 1, the heavy chain CDR2 sequence is shown in SEQID NO: 2, and the heavy chain CDR3 sequence is shown in SEQID NO: 3; the light chain CDR1 sequence is shown in SEQID NO: 4, the light chain CDR2 sequence is shown in SEQID NO: 5, and the light chain CDR3 sequence is shown in SEQID NO:

6.

2. The thyroid stimulating hormone monoclonal antibody 8I22 according to claim 1, characterized in that The heavy chain variable region sequence of the monoclonal antibody 8I22 is shown in SEQ ID NO: 7, and the light chain variable region sequence is shown in SEQ ID NO:

8.

3. The thyroid stimulating hormone monoclonal antibody 8I22 according to claim 1, characterized in that The heavy chain sequence of the monoclonal antibody 8I22 is shown in SEQ ID NO: 9, and the light chain sequence is shown in SEQ ID NO:

10.

4. A thyroid stimulating hormone monoclonal antibody 9E6, characterized in that The heavy chain CDR1 sequence of the monoclonal antibody 9E6 is shown in SEQID NO:11, the heavy chain CDR2 sequence is shown in SEQID NO:12, and the heavy chain CDR3 sequence is shown in SEQID NO:13; the light chain CDR1 sequence is shown in SEQID NO:14, the light chain CDR2 sequence is shown in SEQID NO:15, and the light chain CDR3 sequence is shown in SEQID NO:

16.

5. The thyroid stimulating hormone monoclonal antibody 9E6 according to claim 4, characterized in that The heavy chain variable region sequence of the monoclonal antibody 9E6 is shown in SEQ ID NO: 17, and the light chain variable region sequence is shown in SEQ ID NO:

18.

6. The thyroid stimulating hormone monoclonal antibody 9E6 according to claim 1, characterized in that The heavy chain sequence of the monoclonal antibody 9E6 is shown in SEQ ID NO: 19, and the light chain sequence is shown in SEQ ID NO:

20.

7. A fluorescent immunochromatographic detection agent for thyroid stimulating hormone, characterized in that: include: TSH fluorescent immunochromatographic detection microspheres and capture antibodies; The TSH fluorescent immunochromatographic detection microspheres are prepared by first preparing avidin-labeled fluorescent microspheres; labeling the monoclonal antibody 9E6 according to any one of claims 4 to 6 with biotin to obtain biotin-labeled monoclonal antibody 9E6; and then mixing the avidin-labeled fluorescent microspheres and the biotin-labeled monoclonal antibody 9E6 to obtain the TSH fluorescent immunochromatographic detection microspheres. The capture antibody is the monoclonal antibody 8I22 according to any one of claims 1 to 3.

8. A fluorescent immunochromatographic detection card for thyroid stimulating hormone, characterized in that: A fluorescent immunochromatographic detection agent for thyroid stimulating hormone according to claim 7.

9. The fluorescent immunochromatographic detection reagent card for thyroid stimulating hormone according to claim 8, characterized in that: Includes absorbent paper, nitrocellulose membrane, conjugate pad and blank substrate; The TSH fluorescent immunochromatographic detection microspheres are provided on the conjugate pad; The nitrocellulose membrane is coated with a detection T line and a quality control C line, wherein the detection T line is a capture antibody and the quality control C line is an avidin polyclonal antibody.

10. The fluorescent immunochromatographic detection card for thyroid stimulating hormone according to claim 9, characterized in that: The coating amount of the detection T line is 0.5~1.0 mg / mL; the coating amount of the quality control C line is 0.2~0.5 mg / mL.

Images