Anti-methylenedioxymethamphetamine antibodies, reagents and kits for detecting methylenedioxymethamphetamine

Abstract

The invention discloses an anti-methylenedioxymethamphetamine antibody, a reagent and a kit for detecting methylenedioxymethamphetamine, and relates to the technical field of antibodies. The anti-methylenedioxymethamphetamine antibody disclosed in the invention comprises a heavy chain complementary determining region and a light chain complementary determining region. The antibody provides an important source of raw materials for the detection of methylenedioxymethamphetamine, and has improved affinity and activity.

Description

Technical Field

[0001] The present invention relates to the technical field of antibodies, and in particular to an anti-methylenedioxymethamphetamine antibody, a reagent and a kit for detecting methylenedioxymethamphetamine. Background Art

[0002] Methylenedioxymethamphetamine (MDMA), scientific name: N, a-dimethyl-3-4-methylenedioxyphenylethylamine, molecular formula C11H15O2N, has a strong hallucinogenic effect. Long-term abuse of MDMA will easily lead to manic and angry temper, violent tendencies, and induce the occurrence of vicious violent cases. There will also be mental symptoms such as depression, anxiety, increased heart rate, increased blood pressure, or other cognitive impairment and even sudden death. Because of its addictive and dependent nature, the relevant national departments have listed it as a drug.

[0003] There have been reports on the detection methods of MDMA, such as gas chromatography, gas chromatography / mass spectrometry, and high-performance liquid chromatography. High-performance capillary electrophoresis is an analytical method developed in recent years. It has the characteristics of strong separation ability, high separation efficiency, short analysis time, and small amount of sample and solvent required. Micellar capillary electrokinetic chromatography has been used abroad, but it takes a long time, so a fast and convenient detection method is needed.

[0004] The principle of colloidal gold test paper is to use the specific binding reaction of antigen and antibody and immunochromatography technology, and use a specific color reaction to accurately display the test results on the test paper. It has the characteristics of high sensitivity, strong specificity, good repeatability, high accuracy, simple operation, and rapid and accurate detection. The key to preparing colloidal gold test paper is to obtain antibodies against MDMA. Therefore, there is a strong demand in this field for antibodies that effectively bind to MDMA and detect it.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The present application provides an anti-MDA antibody with improved affinity and / or activity to improve the detection of MDA and provide an important source of raw materials for the detection of MDA.

[0007] In order to achieve the above-mentioned object, according to one aspect of the present invention, an anti-methylenedioxymethamphetamine antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises a HCDR1 having an amino acid sequence as shown in SEQ ID NO:1 or 15, a HCDR2 as shown in SEQ ID NO:2, a HCDR3 as shown in SEQ ID NO:3, and an LCDR1 having an amino acid sequence as shown in any one of SEQ ID NO:4, 16, 17, and 18, a LCDR2 as shown in SEQ ID NO:5, and a LCDR3 as shown in SEQ ID NO:6.

[0008] In order to achieve the above object, according to the second aspect of the present invention, an anti-methylenedioxymethamphetamine antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises a heavy chain variable region and / or a light chain variable region, the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, the light chain variable region comprises a sequence structure of LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4, and the amino acid sequences of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are the amino acid sequences of the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0009] In order to achieve the above object, according to the third aspect of the present invention, an anti-methylenedioxymethamphetamine antibody is provided, comprising a heavy chain and / or a light chain, wherein the heavy chain comprises the above-mentioned heavy chain variable region, and the light chain comprises the above-mentioned light chain variable region.

[0010] In order to achieve the above object, according to the fourth aspect of the present invention, an antibody conjugate is provided, wherein the antibody conjugate comprises the above antibody or a functional fragment thereof.

[0011] In order to achieve the above object, according to the fifth aspect of the present invention, a reagent or kit for detecting methylenedioxymethamphetamine is provided, wherein the reagent or kit comprises the above-mentioned antibody or its functional fragment or the above-mentioned antibody conjugate.

[0012] In order to achieve the above-mentioned object, according to the sixth aspect of the present invention, a method for detecting methylenedioxymethamphetamine is provided, comprising: contacting the above-mentioned antibody or its functional fragment, conjugate, reagent or kit with methylenedioxymethamphetamine in a sample to be detected to form an immune complex.

[0013] In order to achieve the above object, the present invention also provides a vector, a cell and a method for preparing the above antibody or its functional fragment. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0015] Figure 1 The figure shows the results of reducing SDS-PAGE of Anti-MDMA 8C6mut1 to mut6. DETAILED DESCRIPTION

[0016] The present invention provides an anti-methylenedioxymethamphetamine antibody or a functional fragment thereof, wherein the antibody or the functional fragment thereof comprises an amino acid sequence such as HCDR1 shown in SEQ ID NO: 1 or 15, HCDR2 shown in SEQ ID NO: 2, HCDR3 shown in SEQ ID NO: 3, and an amino acid sequence such as LCDR1 shown in any one of SEQ ID NO: 4, 16, 17, and 18, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6. The above-mentioned antibody has improved affinity and / or activity.

[0017] In the present invention, the term "antibody" is used in the broadest sense and may include full-length monoclonal antibodies, bispecific or multispecific antibodies, and chimeric antibodies, so long as they exhibit the desired biological activity.

[0018] In the present invention, the term "complementarity determining region", "CDR" or "CDRs" refers to the highly variable region of the heavy and light chains of immunoglobulins, and refers to the region containing one or more or even all of the major amino acid residues that play a role in the binding of an antibody or antigen-binding fragment to its recognized antigen or epitope. In a specific embodiment of the present invention, CDRs refers to the highly variable region of the heavy and light chains of the antibody.

[0019] In the present invention, the heavy chain complementary determining region is represented by HCDR, which includes HCDR1, HCDR2 and HCDR3; the light chain complementary determining region is represented by LCDR, which includes LCDR1, LCDR2 and LCDR3. Commonly used CDR labeling methods in the art include: Kabat numbering scheme, IMGT numbering scheme, Chothia and Lesk numbering scheme, and the new standardized numbering system introduced by Lefranc et al. for all protein sequences of the immunoglobulin superfamily in 1997. Kabat et al. were the first to propose a standardized numbering scheme for immunoglobulin variable regions. Over the past few decades, the accumulation of sequences has led to the creation of the KABATMAN database, and the Kabat numbering scheme is generally considered to be a widely used standard for numbering antibody residues. The present invention uses the Kabat annotation standard to mark the CDR region, but CDR regions marked by other methods also fall within the scope of protection of the present invention.

[0020] In the present invention, the "framework region" or "FR" region includes the heavy chain framework region and the light chain framework region, and refers to the region other than the CDR in the heavy chain variable region and the light chain variable region of the antibody; wherein the heavy chain framework region can be further subdivided into adjacent regions separated by CDR, including HFR1, HFR2, HFR3 and HFR4 framework regions; the light chain framework region can be further subdivided into adjacent regions separated by CDR, including LFR1, LFR2, LFR3 and LFR4 framework regions.

[0021] In the present invention, the heavy chain variable region is obtained by arranging and connecting the following CDRs and FRs in the following combinations: HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4; the light chain variable region is obtained by arranging and connecting the following CDRs and FRs in the following combinations: LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4.

[0022] In an optional embodiment, the antibody or its functional fragment further comprises HFR1, HFR2, HFR3, HFR4 having amino acid sequences as shown in SEQ ID NO:7 to SEQ ID NO:10 and LFR1, LFR2, LFR3 and LFR4 having amino acid sequences as shown in SEQ ID NO:11 to SEQ ID NO:14.

[0023] It should be noted that, in other embodiments, the amino acid sequences of the framework regions of the antibodies or functional fragments thereof provided by the present invention may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned corresponding framework regions (SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14).

[0024] Optionally, the amino acid sequence of LFR1 of the antibody or its functional fragment is as shown in SEQ ID NO: 19;

[0025] Optionally, the antibody or its functional fragment has a KD of less than 7.34×10 -9 The affinity of M for binding to MDMA.

[0026] In an optional embodiment, the antibody or its functional fragment has a KD≤10 -9 M, KD≤10 -10 M, KD≤10 -11 M or KD≤10 -12 Affinity of M for binding to MDMA.

[0027] In an optional embodiment, the antibody or its functional fragment has a KD≤2.83×10 -10 Affinity of M for binding to MDMA.

[0028] In an optional embodiment, the antibody or its functional fragment has a KD≤1.0×10 -12 Affinity of M for binding to MDMA.

[0029] On the other hand, an embodiment of the present invention provides an anti-methylenedioxymethamphetamine antibody or a functional fragment thereof, wherein the antibody or the functional fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, and the light chain variable region comprises a sequence structure of LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4, and the amino acid sequences of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are the above-mentioned amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, and the amino acid sequences of the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 are the above-mentioned amino acid sequences of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4.

[0030] In an optional embodiment, the heavy chain variable region amino acid sequence is shown in any one of SEQ ID NOs: 22 to 23.

[0031] In an optional embodiment, the light chain variable region amino acid sequence is shown in any one of SEQ ID NOs: 24 to 31.

[0032] In an alternative embodiment, the antibody further comprises a constant region.

[0033] In an alternative embodiment, the constant region includes a heavy chain constant region and / or a light chain constant region.

[0034] In an alternative embodiment, the heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD, and the light chain constant region is selected from the kappa or lambda type light chain constant region.

[0035] In an alternative embodiment, the constant region is derived from a species of cow, horse, dairy cow, pig, sheep, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose, turkey, fighting cock or human.

[0036] In an alternative embodiment, the species origin of the constant region is mouse.

[0037] In an optional embodiment, the heavy chain constant region sequence (CH) is as shown in SEQ ID NO:20, and the light chain constant region (CL) sequence is as shown in SEQ ID NO:21.

[0038] It should be noted that, in other embodiments, the constant region sequence may have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the above-mentioned constant region (SEQ ID NO: 18 or 19).

[0039] In an optional embodiment, the functional fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

[0040] The functional fragments of the above antibodies usually have the same binding specificity as the antibodies from which they are derived. It is easy for a person skilled in the art to understand based on the contents described in the present invention that the functional fragments of the above antibodies can be obtained by methods such as enzymatic digestion (including pepsin or papain) and / or by chemical reduction to split disulfide bonds. Based on the structure of the complete antibody disclosed in the present invention, a person skilled in the art can easily obtain the above functional fragments.

[0041] The functional fragments of the above antibodies can also be synthesized by recombinant genetic techniques known to those skilled in the art or by, for example, an automatic peptide synthesizer, such as those sold by Applied BioSystems and the like.

[0042] On the other hand, the present invention provides an anti-methylenedioxymethamphetamine antibody, comprising a heavy chain and / or a light chain, wherein the heavy chain comprises the above-mentioned heavy chain variable region and the above-mentioned heavy chain constant region; and the light chain comprises the above-mentioned light chain variable region and the above-mentioned light chain constant region.

[0043] In an optional embodiment, the amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 32 to 33.

[0044] In an optional embodiment, the amino acid sequence of the light chain is as shown in any one of SEQ ID NO:34 to 41.

[0045] In another aspect, the present invention provides an antibody conjugate, comprising the above-mentioned antibody, wherein the antibody is directly or indirectly covalently coupled to the object to be coupled, or the antibody is coupled to the object to be coupled by non-covalent adsorption.

[0046] In an alternative embodiment, the antibody in the above antibody conjugate is conjugated to biotin or a biotin derivative.

[0047] In an alternative embodiment, the antibody in the above antibody conjugate is conjugated to a label.

[0048] In an optional embodiment, the above-mentioned marker refers to a class of substances with properties that can be directly observed by the naked eye or detected or detected by an instrument, such as luminescence, color development, radioactivity, etc., through which qualitative or quantitative detection of the corresponding target can be achieved.

[0049] In an alternative embodiment, the label includes but is not limited to fluorescent dyes, enzymes, radioactive isotopes, chemiluminescent agents and nanoparticle labels.

[0050] In actual use, those skilled in the art can select a suitable marker according to the detection conditions or actual needs. No matter which marker is used, it belongs to the protection scope of the present invention.

[0051] In an optional embodiment, the fluorescent dye is not limited to fluorescein dyes and their derivatives (for example, including but not limited to fluorescein isothiocyanate (FITC), hydroxyfluorescein (FAM), tetrachlorofluorescein (TET), etc. or their analogs), rhodamine dyes and their derivatives (for example, including but not limited to red rhodamine (RBITC), tetramethylrhodamine (TAMRA), rhodamine B (TRITC) or its analogs), Cy series dyes and their derivatives (for example, including but not limited to Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy3, etc. or their analogs), Alexa series dyes and their derivatives (for example, including but not limited to AlexaFluor350, 405, 430, 488, 532, 546, 555, 568, 594, 610, 33, 647, 680, 700, 750, etc. or their analogs) and protein dyes and their derivatives (for example, including but not limited to phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC), pre-chlorophyll protein (preCP), etc.).

[0052] In alternative embodiments, the enzyme includes, but is not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and 6-phosphoglucose deoxygenase.

[0053] In an optional embodiment, the radioactive isotope includes but is not limited to 212Bi, 131I, 111In, 90Y, 186Re, 211At, 125I, 188Re, 153Sm, 213Bi, 32P, 94mTc, 99mTc, 203Pb, 67Ga, 68Ga, 43Sc, 47Sc, 110mIn, 97Ru, 62Cu, 64Cu, 67Cu, 68Cu, 86Y, 88Y, 121Sn, 161Tb, 166Ho, 105Rh, 177Lu, 172Lu and 18F.

[0054] In an optional embodiment, the chemiluminescent reagent includes but is not limited to luminol and its derivatives, lucigenin, crustacean fluorescein and its derivatives, bipyridine ruthenium and its derivatives, acridinium esters and their derivatives, dioxetanes and their derivatives, lophanes and their derivatives, and peroxyoxalates and their derivatives.

[0055] In an optional embodiment, the nanoparticle markers include, but are not limited to, nanoparticles, colloids, organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles, and rare earth complex nanoparticles.

[0056] In alternative embodiments, the colloid includes, but is not limited to, colloidal metals, disperse dyes, dye-labeled microspheres, and latex.

[0057] In an alternative embodiment, the colloidal metal includes, but is not limited to, colloidal gold, colloidal silver, and colloidal selenium.

[0058] In an optional embodiment, the colloidal metal is colloidal gold.

[0059] In an alternative embodiment, the antibody in the above antibody conjugate is coupled to a solid phase.

[0060] In alternative embodiments, the solid phase is selected from the group consisting of microspheres, plates, and membranes.

[0061] In an optional embodiment, the solid phase includes, but is not limited to, magnetic microspheres, plastic microspheres, plastic microparticles, microplates, glass, capillaries, nylon, and nitrocellulose membranes.

[0062] In an alternative embodiment, the solid phase is a nitrocellulose membrane.

[0063] On the other hand, the present invention provides a reagent or kit for detecting methylenedioxymethamphetamine, wherein the reagent or kit comprises the above-mentioned antibody or its functional fragment or the above-mentioned antibody conjugate.

[0064] On the other hand, the present invention provides use of the above-mentioned antibody or its functional fragment, antibody conjugate or the above-mentioned reagent or kit in the detection of methylenedioxymethamphetamine.

[0065] On the other hand, the present invention provides a method for detecting methylenedioxymethamphetamine, comprising: contacting the above-mentioned antibody or its functional fragment, conjugate, or reagent or kit with methylenedioxymethamphetamine in a sample to be detected to form an immune complex.

[0066] In an optional embodiment, the immune complex further comprises a second antibody, which binds to the antibody or a functional fragment thereof.

[0067] In an optional embodiment, the immune complex further comprises a second antibody, which binds to MDMA.

[0068] In another aspect, the present invention provides a nucleic acid molecule encoding the above-mentioned antibody or a functional fragment thereof.

[0069] In another aspect, the present invention provides a vector comprising the above nucleic acid molecule.

[0070] In another aspect, the present invention provides a cell containing the above-mentioned vector.

[0071] In another aspect, the present invention provides a method for preparing an antibody or a functional fragment thereof, comprising: culturing the cell as described above.

[0072] Based on the amino acid sequence of the antibody or its functional fragment disclosed in the present invention, those skilled in the art can easily think of using genetic engineering technology or other technologies (chemical synthesis, recombinant expression) to prepare the antibody or its functional fragment, for example, separating and purifying the antibody or its functional fragment from the culture product of a recombinant cell that can recombinantly express the antibody or its functional fragment as described in any of the above items, which is easy to achieve for those skilled in the art. Based on this, no matter what technology is used to prepare the antibody or its functional fragment of the present invention, it belongs to the protection scope of the present invention.

[0073] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0074] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used for the practice or testing of the preparations or unit doses herein, some methods and materials are now described. Unless otherwise stated, the techniques adopted or considered herein are standard methods. Materials, methods and examples are illustrative and non-restrictive only.

[0075] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry, and immunology, which are within the capabilities of a skilled artisan. This technique is fully explained in the literature, such as Molecular Cloning: A Laboratory Manual, 2nd Edition (Sambrook et al., 1989); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Animal Cell Culture (RI Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JM Miller and MP Calos, eds., 1987); Current Protocols in Molecular Biology (FM Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction (PCR: The Polymerase Chain Reaction) (Academic Press, Inc., 1987). Reaction" (Mullis et al., eds., 1994); and Current Protocols in Immunology (JE Coligan et al., eds., 1991), each of which is expressly incorporated herein by reference.

[0076] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0077] Example 1 Preparation of Anti-MDMA 8C6 Monoclonal Antibody

[0078] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TMRACE cDNA Amplification Kit was purchased from Takara. pMD-18T vector was purchased from Takara. Plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were completed by Invitrogen.

[0079] 1 Construction of recombinant plasmid

[0080] (1) Antibody gene preparation

[0081] mRNA was extracted from a hybridoma cell line secreting monoclonal antibodies against MDMA, and a DNA product was obtained by RT-PCR. The product was inserted into a pMD-18T vector after A addition reaction with rTaq DNA polymerase and transformed into DH5α competent cells. After colonies grew, Heavy Chain and Light Chain gene clones were obtained, and 4 clones each were sent to a gene sequencing company for sequencing.

[0082] (2) Sequence analysis of the variable region gene of Anti-MDMA 8C6 antibody

[0083] The gene sequences obtained by the above sequencing were placed in the Kabat antibody database for analysis, and the VNTI11.5 software was used for analysis to determine that the genes amplified by the heavy chain and light chain primer pairs were correct. Among the gene fragments amplified by the Light Chain, the VL gene sequence was 336bp, and there was a 57bp leader peptide sequence in front of it; in the gene fragments amplified by the Heavy Chain primer pair, the VH gene sequence was 345bp, belonging to the VH1 gene family, and there was a 57bp leader peptide sequence in front of it.

[0084] (3) Construction of recombinant antibody expression plasmid

[0085] pcDNA TM 3.4 vector is a recombinant antibody eukaryotic expression vector constructed, which has introduced multiple cloning restriction sites such as HindIII, BamHI, and EcoRI, and is named pcDNA3.4A expression vector, hereinafter referred to as 3.4A expression vector; according to the sequencing results of the antibody variable region genes in the above pMD-18T, the VL and VH gene-specific primers of the antibody were designed, with HindIII, EcoRI restriction sites and protective bases at both ends, respectively, and the 0.74kb Light Chain gene fragment and the 1.44kb Heavy Chain gene fragment were amplified by the PCR amplification method.

[0086] The Heavy Chain and Light Chain gene fragments were double-digested with HindIII / EcoRI, and the 3.4A vector was double-digested with HindIII / EcoRI. After the fragments and vectors were purified and recovered, the Heavy Chain gene and the Light Chain gene were respectively connected to the 3.4A expression vector to obtain the recombinant expression plasmids of Heavy Chain and Light Chain, respectively.

[0087] 2. Sample preparation of recombinant antibodies

[0088] Resuscitate HEK293 cells in advance, subculture to 200ml system, make the cell density reach 3-5×106cells / ml, cell viability>95%; wash the cells by centrifugation, re-dissolve with culture medium, and adjust the cell density to 2.9×106cells / ml as cell diluent. Prepare plasmid DNA and transfection reagent diluent with culture medium respectively. Add transfection reagent diluent to plasmid DNA diluent, mix well and let stand at room temperature for 15min; slowly add the mixture to cell diluent within 1min, mix well and count, record and observe the viability of cells after transfection, and place them in a 35℃ constant temperature incubator for culture, speed 120rmp, CO2 content 8%, and centrifuge after 13 days. The supernatant of centrifugation was affinity purified using proteinA affinity chromatography column. Take 6μg purified antibody for reducing SDS-PAGE, and the electrophoresis is shown in the figure. After reducing SDS-PAGE, two bands were shown, one with Mr of 50 KD (heavy chain) and the other with Mr of 28 KD (light chain).

[0089] Example 2 Affinity and activity optimization

[0090] Although the Anti-MDMA 8C6 monoclonal antibody obtained in Example 1 has the ability to bind to the methylenedioxymethamphetamine antigen, its affinity and antibody activity are not ideal, so the applicant performs directed mutation on the light chain CDR and heavy chain CDR of the antibody. That is, the antibody variable region structure simulation, antigen and antibody variable region complex structure simulation, antibody key amino acid analysis and mutation design are performed by computer, bidirectional primers covering the mutation site are synthesized according to the mutation scheme, primers at both ends of the target DNA are synthesized, high-fidelity PCR reaction is performed, the PCR product is cloned into a vector, and then the mutant antibody is prepared according to the method described in Example 1. After screening, monoclonal antibodies with significantly improved affinity and antibody activity are obtained, and are named: Anti-MDMA 8C6mut1 to Anti-MDMA 8C6mut6. The heavy chain and light chain amino acid sequences of each monoclonal antibody are shown in the following table respectively.

[0091] Table 1 Antibody sequences

[0092] Sample name Heavy chain number Light chain number Anti-MDMA 8C6mut1 SEQ ID NO:32 SEQ ID NO:34 Anti-MDMA 8C6mut2 SEQ ID NO:32 SEQ ID NO:37 Anti-MDMA 8C6mut3 SEQ ID NO:32 SEQ ID NO:39 Anti-MDMA 8C6mut4 SEQ ID NO:33 SEQ ID NO:34 Anti-MDMA 8C6mut5 SEQ ID NO:32 SEQ ID NO:41 Anti-MDMA 8C6mut6 SEQ ID NO:32 SEQ ID NO:36

[0093] Example 3 Performance Testing of Antibodies

[0094] 1 Affinity analysis

[0095] Using the AMC sensor, the purified antibody was diluted to 10 ug / ml with PBST, and MDMA-BSA (purchased from EastCoast Bio, catalog number P82-99-51A) was graded diluted with PBST.

[0096] Operation process: 60s of equilibrium in buffer 1 (PBST), 300s of antibody fixation in antibody solution, 180s of incubation in buffer 2 (PBST), 420s of binding in antigen solution, 1200s of dissociation in buffer 2, sensor regeneration with 10mM pH 1.69 GLY solution and buffer 3 (PBST), and data output. (KD represents equilibrium dissociation constant, i.e. affinity; kon represents binding rate; kdis represents dissociation rate. The main components of PBST are Na2HPO4+NaCl+TW-20).

[0097] Table 2 Affinity data

[0098]

[0099]

[0100] 2. Activity identification

[0101] Coating solution (main component NaHCO3) diluted MDMA-BSA (purchased from EastCoast Bio, item number P82-99-51A) to 2ug / ml, 100μL per well, 4℃ overnight; the next day, wash twice with washing solution, pat dry; add blocking solution (20% BSA + 80% PBS), 120μl per well, 37℃, 1h, pat dry; add diluted monoclonal antibody, 100μl / well, 37℃, 30min-60min; wash 5 times with washing solution, pat dry; add goat anti-mouse IgG-HRP, 100μl per well, 37℃, 30min; wash 5 times with washing solution (PBS), pat dry; add color development solution A (50μl / well, containing 2.1g / L citric acid, 12.25g / L citric acid, 0.07g / L acetanilide and 0.5g / L urea peroxide), add colorimetric solution B (50μl / well, containing 1.05g / L citric acid, 0.186g / EDTA·2Na, 0.45g / L TMB and 0.2ml / L concentrated HCl), 10min; add stop solution (50μl / well, containing 0.75g / EDTA·2Na and 10.2ml / L concentrated H2SO4); read OD value at 450nm (reference 630nm) on the microplate reader. The reactivity of the expressed recombinant antibody with BSA was detected by the same method as above. The results showed that the self-produced antibody still reacted with BSA at an OD of less than 0.1 when the dosage reached 10ug / ml, indicating that the recombinant antibody expressed this time was inactive to BSA, further proving that the expressed recombinant antibody specifically recognized MDMA. The results are shown in the table below.

[0102] Table 3 Activity data

[0103] Sample concentration (ng / ml) 62.50 31.25 15.63 7.81 3.91 0 Comparison 1.224 0.765 0.401 0.205 0.101 0.032 Anti-MDMA 8C6mut1 1.88 1.12 0.717 0.388 0.301 0.038 Anti-MDMA 8C6mut2 1.732 1.093 0.695 0.326 0.291 0.038 Anti-MDMA 8C6mut3 1.663 1.074 0.571 0.261 0.108 0.044 Anti-MDMA 8C6mut4 1.612 1.034 0.598 0.231 0.106 0.031 Anti-MDMA 8C6mut5 1.863 1.174 0.671 0.381 0.268 0.044 Anti-MDMA 8C6mut6 1.966 1.402 0.845 0.499 0.335 0.033

[0104] 3. Stability Assessment

[0105] The above antibodies were placed at 4°C (refrigerator), -80°C (refrigerator), and 37°C (constant temperature box) for 21 days, and samples were taken for 7 days, 14 days, and 21 days for status observation, and the activity of the 21-day samples was tested. The results showed that there was no obvious protein state change in the antibodies under the three test conditions for 21 days, and the activity did not show a downward trend with the increase of the test temperature, indicating that the above antibodies are stable. The following table shows the OD results of the enzyme immunoassay activity test for 21 days of antibody test.

[0106] Table 4 Stability data

[0107]

[0108]

[0109] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

[0110] The partial amino acid sequences involved in this application are as follows:

[0111] Sequence number Sequence fragment SEQ ID NO:1 SDIMS SEQ ID NO:2 TISSGGSHTYYPDSVKG SEQ ID NO:3 GDJ SEQ ID NO:4 RSSQNLVHSSGLTHLE SEQ ID NO:5 KVSNRFS SEQ ID NO:6 FQGHA SEQ ID NO:15 SDLMS SEQ ID NO:16 RSSQNIVHSSGLTHLE SEQ ID NO:17 RSSQNLVHSSGITHLE SEQ ID NO:18 RSSQNIVHSSGITHLE SEQUENCE LISTING <110> Dongguan Pengzhi Biotechnology Co., Ltd. <120> Anti-methylenedioxymethamphetamine antibodies, reagents and kits for detecting methylenedioxymethamphetamine <130> P2022089CN01 <160> 41 <170> PatentIn version 3.5 <210> 1 <211> 5 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 1 Ser Asp Ile Met Ser 1 5 <210> 2 <211> 17 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 2 Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro Asp Ser Val Lys 1 5 10 15 Gly <210> 3 <211> 6 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 3 Gly Asp Tyr Gly Asp Asn 1 5 <210> 4 <211> 16 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 4 Arg Ser Ser Gln Asn Leu Val His Ser Ser Gly Leu Thr His Leu Glu 1 5 10 15 <210> 5 <211> 7 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 5 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 6 <211> 7 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 6 Phe Gln Gly Ala His Ala Pro 1 5 <210> 7 <211> 30 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 7 Asp Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Glu Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Ile Ser 20 25 30 <210> 8 <211> 14 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 8 Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val Ala 1 5 10 <210> 9 <211> 32 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 9 Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Leu Tyr Leu Gln 1 5 10 15 Met Arg Ser Leu Lys Ser Val Asp Thr Ala Met Tyr Tyr Cys Leu Arg 20 25 30 <210> 10 <211> 11 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 10 Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser 1 5 10 <210> 11 <211> twenty three <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 11 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys 20 <210> 12 <211> 15 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 12 Trp Tyr Leu Gln Arg Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr 1 5 10 15 <210> 13 <211> 32 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 13 Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys 20 25 30 <210> 14 <211> 12 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 14 Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 1 5 10 <210> 15 <211> 5 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 15 Asp Leu Met Ser 1 5 <210> 16 <211> 16 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 16 Arg Ser Ser Gln Asn Ile Val His Ser Ser Gly Leu Thr His Leu Glu 1 5 10 15 <210> 17 <211> 16 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 17 Arg Ser Ser Gln Asn Leu Val His Ser Ser Gly Ile Thr His Leu Glu 1 5 10 15 <210> 18 <211> 16 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 18 Arg Ser Ser Gln Asn Ile Val His Ser Ser Gly Ile Thr His Leu Glu 1 5 10 15 <210> 19 <211> 23 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 19 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys 20 <210> 20 <211> 336 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 20 Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly Cys Gly 1 5 10 15 Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr 20 25 30 Phe Pro Glu Ser Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser 35 40 45 Ser Val His Thr Phe Pro Ala Leu Leu Gln Ser Gly Leu Tyr Thr Met 50 55 60 Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val 65 70 75 80 Thr Cys Ser Val Ala His Pro Ala Ser Ser Thr Thr Val Asp Lys Lys 85 90 95 Leu Glu Pro Ser Gly Pro Ile Ser Thr Ile Asn Pro Cys Pro Pro Cys 100 105 110 Lys Glu Cys His Lys Cys Pro Ala Pro Asn Leu Glu Gly Gly Pro Ser 115 120 125 Val Phe Ile Phe Pro Pro Asn Ile Lys Asp Val Leu Met Ile Ser Leu 130 135 140 Thr Pro Lys Val Thr Cys Val Val Val Asp Val Ser Glu Asp Asp Pro 145 150 155 160 Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val His Thr Ala 165 170 175 Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Ile Arg Val Val 180 185 190 Ser Thr Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe 195 200 205 Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ser Pro Ile Glu Arg Thr 210 215 220 Ile Ser Lys Ile Lys Gly Leu Val Arg Ala Pro Gln Val Tyr Ile Leu 225 230 235 240 Pro Pro Pro Ala Glu Gln Leu Ser Arg Lys Asp Val Ser Leu Thr Cys 245 250 255 Leu Val Val Gly Phe Asn Pro Gly Asp Ile Ser Val Glu Trp Thr Ser 260 265 270 Asn Gly His Thr Glu Glu Asn Tyr Lys Asp Thr Ala Pro Val Leu Asp 275 280 285 Ser Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Met Lys Thr Ser 290 295 300 Lys Trp Glu Lys Thr Asp Ser Phe Ser Cys Asn Val Arg His Glu Gly 305 310 315 320 Leu Lys Asn Tyr Tyr Leu Lys Lys Thr Ile Ser Arg Ser Pro Gly Lys 325 330 335 <210> 21 <211> 107 <212> PRT <213> Artificial <220> <223> artificial sequence <400> 21 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 1 5 10 15 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 20 25 30 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 35 40 45 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 65 70 75 80 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 85 90 95 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 100 105 <210> 22 <211> 115 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 22 Asp Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Glu Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Ile Ser Ser Asp 20 25 30 Ile Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Arg Ser Leu Lys Ser Val Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Leu Arg Gly Asp Tyr Gly Asp Asn Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser 115 <210> 23 <211> 115 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 23 Asp Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Glu Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Ile Ser Ser Asp 20 25 30 Leu Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Arg Ser Leu Lys Ser Val Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Leu Arg Gly Asp Tyr Gly Asp Asn Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser 115 <210> 24 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 24 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 25 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 25 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 26 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 26 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 27 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 27 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 28 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 28 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 29 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 29 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 30 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 30 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 31 <211> 112 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 31 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 32 <211> 451 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 32 Asp Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Glu Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Ile Ser Ser Asp 20 25 30 Ile Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Arg Ser Leu Lys Ser Val Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Leu Arg Gly Asp Tyr Gly Asp Asn Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro 115 120 125 Gly Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val 130 135 140 Lys Gly Tyr Phe Pro Glu Ser Val Thr Val Thr Trp Asn Ser Gly Ser 145 150 155 160 Leu Ser Ser Ser Val His Thr Phe Pro Ala Leu Leu Gln Ser Gly Leu 165 170 175 Tyr Thr Met Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser 180 185 190 Gln Thr Val Thr Cys Ser Val Ala His Pro Ala Ser Ser Thr Thr Val 195 200 205 Asp Lys Lys Leu Glu Pro Ser Gly Pro Ile Ser Thr Ile Asn Pro Cys 210 215 220 Pro Pro Cys Lys Glu Cys His Lys Cys Pro Ala Pro Asn Leu Glu Gly 225 230 235 240 Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Ile Lys Asp Val Leu Met 245 250 255 Ile Ser Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Val Ser Glu 260 265 270 Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val 275 280 285 His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Ile 290 295 300 Arg Val Val Ser Thr Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly 305 310 315 320 Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ser Pro Ile 325 330 335 Glu Arg Thr Ile Ser Lys Ile Lys Gly Leu Val Arg Ala Pro Gln Val 340 345 350 Tyr Ile Leu Pro Pro Pro Ala Glu Gln Leu Ser Arg Lys Asp Val Ser 355 360 365 Leu Thr Cys Leu Val Val Gly Phe Asn Pro Gly Asp Ile Ser Val Glu 370 375 380 Trp Thr Ser Asn Gly His Thr Glu Glu Asn Tyr Lys Asp Thr Ala Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Met 405 410 415 Lys Thr Ser Lys Trp Glu Lys Thr Asp Ser Phe Ser Cys Asn Val Arg 420 425 430 His Glu Gly Leu Lys Asn Tyr Tyr Leu Lys Lys Thr Ile Ser Arg Ser 435 440 445 Pro Gly Lys 450 <210> 33 <211> 451 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 33 Asp Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Glu Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Ile Ser Ser Asp 20 25 30 Leu Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Arg Ser Leu Lys Ser Val Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Leu Arg Gly Asp Tyr Gly Asp Asn Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro 115 120 125 Gly Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val 130 135 140 Lys Gly Tyr Phe Pro Glu Ser Val Thr Val Thr Trp Asn Ser Gly Ser 145 150 155 160 Leu Ser Ser Ser Val His Thr Phe Pro Ala Leu Leu Gln Ser Gly Leu 165 170 175 Tyr Thr Met Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser 180 185 190 Gln Thr Val Thr Cys Ser Val Ala His Pro Ala Ser Ser Thr Thr Val 195 200 205 Asp Lys Lys Leu Glu Pro Ser Gly Pro Ile Ser Thr Ile Asn Pro Cys 210 215 220 Pro Pro Cys Lys Glu Cys His Lys Cys Pro Ala Pro Asn Leu Glu Gly 225 230 235 240 Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Ile Lys Asp Val Leu Met 245 250 255 Ile Ser Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Val Ser Glu 260 265 270 Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val 275 280 285 His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Ile 290 295 300 Arg Val Val Ser Thr Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly 305 310 315 320 Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ser Pro Ile 325 330 335 Glu Arg Thr Ile Ser Lys Ile Lys Gly Leu Val Arg Ala Pro Gln Val 340 345 350 Tyr Ile Leu Pro Pro Pro Ala Glu Gln Leu Ser Arg Lys Asp Val Ser 355 360 365 Leu Thr Cys Leu Val Val Gly Phe Asn Pro Gly Asp Ile Ser Val Glu 370 375 380 Trp Thr Ser Asn Gly His Thr Glu Glu Asn Tyr Lys Asp Thr Ala Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Met 405 410 415 Lys Thr Ser Lys Trp Glu Lys Thr Asp Ser Phe Ser Cys Asn Val Arg 420 425 430 His Glu Gly Leu Lys Asn Tyr Tyr Leu Lys Lys Thr Ile Ser Arg Ser 435 440 445 Pro Gly Light 450 <210> 34 <211> 219 <212> PRT <213> Artificial <220> <223> artificial sequence <400> 34 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 35 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 35 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 36 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 36 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 37 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 37 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 38 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 38 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 39 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial Sequence <400> 39 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Ile Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 40 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 40 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 41 <211> 219 <212> PRT <213> Artificial <220> <223> Artificial sequence <400> 41 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Leu Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Ser Gly Leu Thr His Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Tyr Cys Phe Gln Gly 85 90 95 Ala His Ala Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215

Claims

1. An anti-methylenedioxymethamphetamine antibody or an antigen-binding fragment thereof, characterized in that: The antibody or antigen-binding fragment thereof comprises: The amino acid sequence of HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, and the amino acid sequence of LCDR1 shown in SEQ ID NO:4, LCDR2 shown in SEQ ID NO:5, LCDR3 shown in SEQ ID NO:6; or The amino acid sequence of HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, and the amino acid sequence of LCDR1 shown in SEQ ID NO:16, LCDR2 shown in SEQ ID NO:5, LCDR3 shown in SEQ ID NO:6; or The amino acid sequence of HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, and the amino acid sequence of LCDR1 shown in SEQ ID NO:18, LCDR2 shown in SEQ ID NO:5, LCDR3 shown in SEQ ID NO:6; or The amino acid sequence of HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, and the amino acid sequence of LCDR1 shown in SEQ ID NO:17, LCDR2 shown in SEQ ID NO:5, LCDR3 shown in SEQ ID NO:6; or The amino acid sequence is HCDR1 shown in SEQ ID NO: 15, HCDR2 shown in SEQ ID NO: 2, HCDR3 shown in SEQ ID NO: 3, and the amino acid sequence is LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, LCDR3 shown in SEQ ID NO:

6.

2. The antibody or antigen-binding fragment thereof according to claim 1, characterized in that: The antibody or antigen-binding fragment thereof also includes HFR1, HFR2, HFR3, HFR4 whose amino acid sequences are shown in SEQ ID NO:7 to SEQ ID NO:10, and LFR1, LFR2, LFR3 and LFR4 whose amino acid sequences are shown in SEQ ID NO:11 to SEQ ID NO:14, or amino acid sequences that are at least 80% identical to the sequences.

3. The antibody or antigen-binding fragment thereof according to claim 2, characterized in that: The amino acid sequence of LFR1 of the antibody or antigen-binding fragment thereof is shown in SEQ ID NO:

19.

4. The antibody or antigen-binding fragment thereof according to claim 1, characterized in that: The antibody or antigen-binding fragment thereof has a KD of less than 7.34×10 -9 Affinity binding of M against MDMA.

5. An anti-methylenedioxymethamphetamine antibody or an antigen-binding fragment thereof, characterized in that: The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, the light chain variable region comprises a sequence structure of LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4, the amino acid sequence of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3 is the amino acid sequence of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3 described in claim 1, and the amino acid sequence of the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, LFR4 is the amino acid sequence of the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, LFR4 described in claim 2.

6. An anti-methylenedioxymethamphetamine antibody or an antigen-binding fragment thereof, characterized in that: The antibody or antigen-binding fragment thereof comprises a heavy chain variable region as shown in SEQ ID NO: 22 and a light chain variable region as shown in SEQ ID NO: 24; or The heavy chain variable region shown in SEQ ID NO: 22, the light chain variable region shown in SEQ ID NO: 26; or The heavy chain variable region shown in SEQ ID NO:22, the light chain variable region shown in SEQ ID NO:27; or The heavy chain variable region shown in SEQ ID NO:22, the light chain variable region shown in SEQ ID NO:29; or The heavy chain variable region shown in SEQ ID NO:22, the light chain variable region shown in SEQ ID NO:31; or The heavy chain variable region is shown in SEQ ID NO:23, and the light chain variable region is shown in SEQ ID NO:

24.

7. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, characterized in that: The antibody or antigen-binding fragment thereof further comprises a constant region.

8. The antibody or antigen-binding fragment thereof according to claim 7, characterized in that: The species origin of the constant region is cow, horse, pig, sheep, goat, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose or human.

9. The antibody or antigen-binding fragment thereof according to claim 8, characterized in that: The species origin of the constant region is mouse.

10. The antibody or antigen-binding fragment thereof according to claim 9, characterized in that: The constant region includes a heavy chain constant region and / or a light chain constant region.

11. The antibody or antigen-binding fragment thereof according to claim 10, characterized in that: The heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD; the light chain constant region is selected from the κ type or λ type light chain constant region.

12. The antibody or antigen-binding fragment thereof according to claim 11, characterized in that: The heavy chain constant region sequence is as shown in SEQ ID NO: 20 or has at least 80% identity thereto; the light chain constant region sequence is as shown in SEQ ID NO: 21 or has at least 80% identity thereto.

13. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 and 8 to 12, characterized in that: The antigen-binding fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

14. An anti-methylenedioxymethamphetamine antibody comprising a heavy chain and a light chain, characterized in that: The heavy chain comprises the heavy chain variable region of claim 6 and the heavy chain constant region of claim 12; the light chain comprises the light chain variable region of claim 6 and the light chain constant region of claim 12.

15. The anti-MDA antibody according to claim 14, characterized in that: comprising a heavy chain as shown in SEQ ID NO: 32 and a light chain as shown in SEQ ID NO: 34; or The heavy chain shown in SEQ ID NO:32, the light chain shown in SEQ ID NO:36; or The heavy chain shown in SEQ ID NO:32, the light chain shown in SEQ ID NO:37; or The heavy chain shown in SEQ ID NO:32, the light chain shown in SEQ ID NO:39; or The heavy chain as shown in SEQ ID NO:32, the light chain as shown in SEQ ID NO:41; or the heavy chain as shown in SEQ ID NO:33, the light chain as shown in SEQ ID NO:

34.

16. An antibody conjugate, characterized in that: The antibody conjugate comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 15.

17. The antibody conjugate according to claim 16, characterized in that The antibody is conjugated to biotin or a biotin derivative.

18. The antibody conjugate according to claim 16, characterized in that The antibody is coupled to a solid phase.

19. The antibody conjugate according to claim 16, characterized in that: The antibody is conjugated to a label.

20. The antibody conjugate according to claim 19, characterized in that The label is selected from fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents and nanoparticle labels.

21. The antibody conjugate according to claim 19, characterized in that The label is colloidal gold.

22. A reagent or kit for detecting methylenedioxymethamphetamine, characterized in that: The reagent or kit comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 15 or the antibody conjugate according to any one of claims 16 to 21.

23. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 15, or the conjugate according to any one of claims 16 to 21, or the reagent or kit according to claim 22 in the preparation of a product for detecting methylenedioxymethamphetamine, characterized in that: include: The antibody or antigen-binding fragment thereof according to any one of claims 1 to 15, the antibody conjugate according to any one of claims 16 to 21, or the reagent or kit according to claim 22 is contacted with methylenedioxymethamphetamine in a sample to be detected to form an immune complex.

24. The use according to claim 23, characterized in that The immune complex also includes a second antibody that binds to the antibody or antigen-binding fragment thereof.

25. The use according to claim 23, characterized in that The immune complex also includes a second antibody that binds to MDMA.

26. A nucleic acid, characterized in that It encodes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 15.

27. A carrier, characterized in that It contains the nucleic acid according to claim 26.

28. A cell, characterized in that It contains the nucleic acid according to claim 26 or the vector according to claim 27.

29. A method for preparing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 15, characterized in that: It includes: Cultivate the cell of claim 28.

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