CN116478060A - Dezocine hapten and artificial antigen as well as preparation methods and applications thereof - Google Patents
Dezocine hapten and artificial antigen as well as preparation methods and applications thereof Download PDFInfo
- Publication number
- CN116478060A CN116478060A CN202310210890.8A CN202310210890A CN116478060A CN 116478060 A CN116478060 A CN 116478060A CN 202310210890 A CN202310210890 A CN 202310210890A CN 116478060 A CN116478060 A CN 116478060A
- Authority
- CN
- China
- Prior art keywords
- dezocine
- hapten
- xin
- artificial
- artificial antigen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- VTMVHDZWSFQSQP-VBNZEHGJSA-N dezocine Chemical compound C1CCCC[C@H]2CC3=CC=C(O)C=C3[C@]1(C)[C@H]2N VTMVHDZWSFQSQP-VBNZEHGJSA-N 0.000 title claims abstract description 126
- 229960003461 dezocine Drugs 0.000 title claims abstract description 123
- 239000000427 antigen Substances 0.000 title claims abstract description 30
- 102000036639 antigens Human genes 0.000 title claims abstract description 30
- 108091007433 antigens Proteins 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 238000001514 detection method Methods 0.000 claims abstract description 49
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 46
- 238000012360 testing method Methods 0.000 claims abstract description 43
- 239000010931 gold Substances 0.000 claims abstract description 34
- 229910052737 gold Inorganic materials 0.000 claims abstract description 34
- 238000000034 method Methods 0.000 claims abstract description 14
- 239000002994 raw material Substances 0.000 claims abstract description 12
- 241001465754 Metazoa Species 0.000 claims abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 29
- 238000006243 chemical reaction Methods 0.000 claims description 22
- QAEDZJGFFMLHHQ-UHFFFAOYSA-N trifluoroacetic anhydride Chemical compound FC(F)(F)C(=O)OC(=O)C(F)(F)F QAEDZJGFFMLHHQ-UHFFFAOYSA-N 0.000 claims description 12
- 238000003908 quality control method Methods 0.000 claims description 11
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 10
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 10
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 claims description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 9
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 238000004809 thin layer chromatography Methods 0.000 claims description 6
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 5
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 5
- 229940098773 bovine serum albumin Drugs 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 229940014800 succinic anhydride Drugs 0.000 claims description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 5
- 150000001718 carbodiimides Chemical class 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 3
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 3
- OMOVVBIIQSXZSZ-UHFFFAOYSA-N [6-(4-acetyloxy-5,9a-dimethyl-2,7-dioxo-4,5a,6,9-tetrahydro-3h-pyrano[3,4-b]oxepin-5-yl)-5-formyloxy-3-(furan-3-yl)-3a-methyl-7-methylidene-1a,2,3,4,5,6-hexahydroindeno[1,7a-b]oxiren-4-yl] 2-hydroxy-3-methylpentanoate Chemical compound CC12C(OC(=O)C(O)C(C)CC)C(OC=O)C(C3(C)C(CC(=O)OC4(C)COC(=O)CC43)OC(C)=O)C(=C)C32OC3CC1C=1C=COC=1 OMOVVBIIQSXZSZ-UHFFFAOYSA-N 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- 230000008878 coupling Effects 0.000 claims description 2
- 238000010168 coupling process Methods 0.000 claims description 2
- 238000005859 coupling reaction Methods 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 230000003053 immunization Effects 0.000 claims description 2
- 238000002649 immunization Methods 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims 1
- 230000008901 benefit Effects 0.000 abstract description 4
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- 239000010839 body fluid Substances 0.000 abstract description 3
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 3
- 230000005847 immunogenicity Effects 0.000 abstract description 3
- 230000009257 reactivity Effects 0.000 abstract description 2
- 239000000123 paper Substances 0.000 description 36
- 210000002700 urine Anatomy 0.000 description 13
- 239000000047 product Substances 0.000 description 12
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- 239000000523 sample Substances 0.000 description 8
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- 241000283973 Oryctolagus cuniculus Species 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 6
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- 235000010703 Modiola caroliniana Nutrition 0.000 description 3
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
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- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 3
- 239000002207 metabolite Substances 0.000 description 3
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- 239000005720 sucrose Substances 0.000 description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 2
- -1 Boc group Chemical group 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000007910 cell fusion Effects 0.000 description 2
- KWGRBVOPPLSCSI-UHFFFAOYSA-N d-ephedrine Natural products CNC(C)C(O)C1=CC=CC=C1 KWGRBVOPPLSCSI-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000003317 immunochromatography Methods 0.000 description 2
- 229960003299 ketamine Drugs 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229960001252 methamphetamine Drugs 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 239000000014 opioid analgesic Substances 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 238000011056 performance test Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 206010013654 Drug abuse Diseases 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-N Gluconic acid Natural products OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- KWGRBVOPPLSCSI-WPRPVWTQSA-N L-Ephedrine Natural products CN[C@@H](C)[C@H](O)C1=CC=CC=C1 KWGRBVOPPLSCSI-WPRPVWTQSA-N 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
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- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 229940035674 anesthetics Drugs 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 239000000935 antidepressant agent Substances 0.000 description 1
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
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- 238000005886 esterification reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- PJMPHNIQZUBGLI-UHFFFAOYSA-N fentanyl Chemical compound C=1C=CC=CC=1N(C(=O)CC)C(CC1)CCN1CCC1=CC=CC=C1 PJMPHNIQZUBGLI-UHFFFAOYSA-N 0.000 description 1
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- 229960003923 gatifloxacin Drugs 0.000 description 1
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- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 102000048260 kappa Opioid Receptors Human genes 0.000 description 1
- 239000002632 kappa opiate receptor agonist Substances 0.000 description 1
- 238000002690 local anesthesia Methods 0.000 description 1
- 239000003589 local anesthetic agent Substances 0.000 description 1
- 229960005015 local anesthetics Drugs 0.000 description 1
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- 230000004630 mental health Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 239000002623 mu opiate receptor antagonist Substances 0.000 description 1
- 229940124636 opioid drug Drugs 0.000 description 1
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- 230000001376 precipitating effect Effects 0.000 description 1
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- 238000007789 sealing Methods 0.000 description 1
- 230000001624 sedative effect Effects 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 208000011117 substance-related disease Diseases 0.000 description 1
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- TVYLLZQTGLZFBW-GOEBONIOSA-N tramadol Natural products COC1=CC=CC([C@@]2(O)[C@@H](CCCC2)CN(C)C)=C1 TVYLLZQTGLZFBW-GOEBONIOSA-N 0.000 description 1
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 1
- 108020001588 κ-opioid receptors Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C233/00—Carboxylic acid amides
- C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
- C07C233/23—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a ring other than a six-membered aromatic ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/948—Sedatives, e.g. cannabinoids, barbiturates
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/56—Ring systems containing bridged rings
- C07C2603/58—Ring systems containing bridged rings containing three rings
- C07C2603/76—Ring systems containing bridged rings containing three rings containing at least one ring with more than six ring members
- C07C2603/80—Ring systems containing bridged rings containing three rings containing at least one ring with more than six ring members containing eight-membered rings
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Biophysics (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Genetics & Genomics (AREA)
- General Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Anesthesiology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a dezocine hapten and an artificial antigen as well as a preparation method and application thereof, wherein dezocine is used as a raw material to synthesize the dezocine artificial hapten and the artificial antigen, the immunogenicity and the reactivity of the obtained antigen are good, corresponding antibodies are easy to obtain when animals are immunized, and the dezocine hapten and the artificial antigen can be used for preparing a dezocine Xin Jiaoti gold detection test paper; the dezocine Xin Jiaoti gold test paper provided by the invention can rapidly and simply detect dezocine in body fluid, has the advantages of low cost, convenience, quickness and the like compared with a high performance liquid chromatography method and the like, can effectively monitor the dezocine dosage, reduces the harm caused by dezocine abuse, and is easy to interpret and suitable for most scenes and users.
Description
Technical Field
The invention belongs to the field of immunological rapid detection, and particularly relates to a dezocine hapten and an artificial antigen as well as a preparation method and application thereof.
Background
Dezocine is a common domestic opioid analgesic, is a kappa receptor agonist and a mu receptor antagonist, mainly excites kappa receptors to produce analgesic and sedative effects, is widely applied to postoperative analgesia at present, has good use effect when being matched with other anesthetics and analgesic drugs, and can effectively reduce pain feeling caused by operation. Besides postoperative analgesia, the anesthesia effect in the operation can be effectively improved through compatibility with local anesthetics. Researches show that dezocine is used as a clinical analgesic, has obvious clinical analgesic effect, is not inferior to equal doses of fentanyl or morphine, and has less adverse reaction caused by injection of dezocine, so that the dezocine can be rapidly popularized and used.
Most dezocine-related studies and reports show that dezocine has smaller addiction, but has less addiction than that of common opioid drugs, but has cases of dezocine Xin Cheng addiction, and patients with repeated use in the cases have a certain dependence on dezocine injection due to Xin Chengyin. Studies show that dezocine has the same action sites as certain antidepressant drugs, can effectively relieve depression of patients, and most addicts represent dezocine to effectively relieve pain, relax body and mind after injection, and help sleep, and these factors promote the patients to dezocine Xin Chengyin. Additionally, the domestic management and control of dezocine is not strict, so that most patients can easily obtain the dezocine by themselves, and the long-term use probability of patients with certain psychological diseases and the probability of addiction are greatly increased.
For the addiction potential of dezocine, on the one hand, the management of dezocine Xin Yaowu needs to be enhanced, and on the other hand, the self-protection consciousness and the correct medication habit of the patient are enhanced. In the management and control of dezote Xin Yaowu, a convenient and accurate detection means is a key part of the management and control.
The immunochromatography colloidal gold technology is widely applied to the fields of medical treatment and production after decades of development, is mature in the field of drug abuse, and has a plurality of drugs and colloidal gold test papers for monitoring drugs on the market. Compared with other detection means, the colloidal gold detection method has the advantages of simpler detection operation, lower detection cost and larger advantage. Dezocine is widely used as an opioid analgesic for postoperative analgesia and local anesthesia, and the national supervision of dezocine does not strictly lead to certain hidden abuse trouble of dezocine. At present, no patent and literature for detecting dezocine abuse-related colloidal gold test paper appears, a certain blank exists for monitoring dezocine abuse by using an immunochromatography colloidal gold technology, and the dezocine abuse monitoring operation is complex and has higher cost by using technologies such as high performance liquid chromatography. The invention mainly innovates and develops a colloidal gold test paper for detecting dezocine and antigens and hapten thereof, provides good detection selection for detecting and controlling abuse of dezocine, helps patients addicted to dezocine Xin Cheng get rid of dependence, and protects physical and mental health of dezocine users.
Disclosure of Invention
A first object of the present invention is to provide a dezocine hapten which addresses the deficiencies of the prior art.
A dezocine hapten has a structural formula shown in a formula (1):
the second purpose of the invention is to provide a preparation method of dezocine hapten, which takes dezocine as raw material, and obtains hapten with carboxyl structure while protecting amino group of dezocine structure through esterification reaction with trifluoroacetic anhydride after protection and deprotection of Boc group; the invention adopts the following technical scheme:
step (1), dezocine is dissolved in tetrahydrofuran, triethylamine and di-tert-butyl dicarbonate are added, and the reaction is carried out for 3 to 5 hours to obtain a product A;
step (2), dissolving the product A in dichloromethane, adding triethylamine, succinic anhydride and 4-Dimethylaminopyridine (DMAP), reacting for 14-16 hours at room temperature, and separating by thin layer chromatography to obtain oily substance B;
step (3), dissolving the oily matter B in a mixed solution of trifluoroacetic acid and dichloromethane, stirring and reacting for 4-6 hours, and evaporating to dryness to obtain oily residue C;
and (4) dissolving oily residue C in benzene, adding trifluoroacetic anhydride for reaction for 1-2 hours at room temperature, then carrying out oil bath reflux reaction for a period of time, and separating by thin layer chromatography to obtain the dezocine artificial hapten.
Preferably, the molar ratio of dezocine, triethylamine and di-tert-butyl dicarbonate in step (1) is (0.5-2): (1.0-3.5): (0.5 to 2.5), more preferably 1:2:1.1.
Preferably, the molar ratio of the product A, triethylamine and succinic anhydride in the step (2) is (0.5-1.5): (1-4): (1-3), more preferably 1:3:2.
Preferably, the molar volume ratio of oil B, trifluoroacetic acid and dichloromethane in step (3) is 0.141mmol:1mL:1mL.
Preferably, in the step (4), the molar ratio of the oily residue C to trifluoroacetic anhydride is (0.8 to 1.3): (1.7 to 2.4), more preferably 1:2.
Preferably, the reflux reaction time in step (4) is 3 hours.
The third object of the invention is to provide a dezocine artificial antigen, which is obtained by coupling and purifying the dezocine hapten with bovine serum albumin, and the structural formula of the dezocine artificial antigen is shown as the formula (2):
the fourth object of the invention is to provide a method for preparing dezocine artificial antigen, which adopts the following technical scheme:
the dezocine artificial hapten, N-hydroxysuccinimide (NHS) and carbodiimide (EDCI) are mixed and then placed into room temperature for reaction for 20 to 24 hours, then added into bovine serum albumin BSA solution for stirring reaction for 3 to 5 hours, and dialyzed in a sodium carbonate solution with the pH value of 10.00 to 13.00 and the mass percent concentration of 0.3 to 0.7 percent and PBS with the concentration of 0.01 to 0.02M in sequence, and the dezocine artificial antigen is obtained after centrifugal precipitation.
Preferably, the molar ratio of the dezocine artificial hapten, the N-hydroxysuccinimide and the carbodiimide is (0.8-1.3): (1.2-1.8): (1.7 to 2.1), more preferably 1:1.5:2.
preferably, the concentration of bovine serum albumin is 4 to 7mg/ml, more preferably 5mg/ml.
The fifth object of the invention is to provide a dezocine Xin Shan clone antibody which is obtained by animal immunization from a dezocine artificial antigen.
The preparation process of the preferred dezote Xin Shan clone antibody of the invention is as follows: the prepared dezocine artificial antigen is purified and diluted, then mixed with an adjuvant for enhancing immunogenicity, injected into the back subcutaneous and thigh muscle of healthy rabbits, repeatedly injected every 3-4 weeks, fresh rabbit blood is collected after 7-9 days of injection each time, the binding titer of the antibody obtained in serum is detected by ELISA method after the rabbit blood is separated, the dezocine Xin Duo clone antibody is obtained by purifying serum after the antibody serum with ideal binding titer is obtained, and then the dezocine Xin Shan clone antibody is obtained by screening and separating and purifying mouse ascites by monoclonal technology by utilizing the B cell fusion tumor cells activated by mouse immunity.
The sixth object of the invention is to provide a Dizodiac Xin Jiaoti gold test paper, which comprises a sample pad, a gold mark pad, a detection line and a quality control line; the detection line is obtained by taking dezote Xin Kangyuan as a raw material spot film.
The preparation process of the spot gold label raw material used for the gold label pad in the dezocine detection test paper is preferably as follows: adding the colloidal gold solution with the pH value adjusted to 9.0 into a proper labeling amount of dezote Xin Shan clone antibody solution, adding a buffer solution after uniformly mixing, centrifuging for 30min at 12000r/min, removing the precipitate, re-suspending with the buffer solution, repeating the centrifugal re-suspending operation for 3 times to obtain stable colloidal gold labeled protein, purifying the labeled protein by using a filtration method, and eluting to obtain the colloidal gold labeled raw material solution for gold labeling.
Preferably, the quality control line raw materials of the Dizodiac Xin Jiaoti gold detection test paper are sheep anti-rabbit IgG and sheep anti-mouse IgG, the concentration in the quality control line spray point solution diluted by phosphate buffer solution is respectively 0.5-2.0 mg/ml and 0.1-0.5 mg/ml, and the mixed solution is uniformly sprayed on an NC film to obtain the quality control line.
According to the invention, preferably, the detection line of the Dizuo Xin Jiaoti gold detection test paper is obtained by uniformly spraying the Dizuo Xin Kangyuan with the concentration of 0.1-2.0 mg/ml serving as a spraying point raw material on an NC film.
The dezocine Xin Jiaoti gold test paper can perform qualitative determination on dezocine and antigens thereof, has higher sensitivity, and the higher the dezocine concentration is, the lower the strength of a detection line is, and the detection limit is 1000ng/mL. The interpretation rule of the dezocine detection test paper in the invention is as follows:
the interpretation is valid: when the quality control line, namely the C line shows a purple red line and the color of the line is obvious, the interpretation of the detection test paper is proved to be effective;
negative: when the quality control line, namely the C line and the detection line, namely the T line, appear mauve and the line color is obvious, the detection sample is proved to have no dezocine and antigen thereof, and the interpretation is negative;
positive: when the quality control line, namely the C line, shows mauve and the line color is obvious, and the detection line, namely the T line, does not show mauve, the detection sample is proved to have dezocine and antigens thereof, and the detection sample is judged to be positive.
The invention has the following beneficial effects:
the dezocine artificial hapten and the artificial antigen are synthesized by taking dezocine as raw materials, the immunogenicity and the reactivity of the obtained antigen are good, corresponding antibodies are easy to obtain when animals are immunized, and the dezocine artificial hapten and the artificial antigen can be used for preparing the dezocine Xin Jiaoti gold detection test paper; the dezocine Xin Jiaoti gold test paper provided by the invention can rapidly and simply detect dezocine in body fluid, has the advantages of low cost, convenience, quickness and the like compared with a high performance liquid chromatography method and the like, can effectively monitor the dezocine dosage, reduces the harm caused by dezocine abuse, and is easy to interpret and suitable for most scenes and users.
Drawings
FIG. 1 is a liquid chromatogram of dezocine artificial hapten I obtained in example 1, wherein mAU represents milliabsorbance units and min represents time units of minutes.
FIG. 2 is an ESI-MS spectrum of dezocine artificial hapten I obtained in example 1, wherein Intans represents signal intensity and m/z represents mass-to-charge ratio.
Detailed Description
The invention will be further analyzed with reference to specific examples.
Example 1: preparation of dezocine artificial hapten
1. 200mg (0.816 mmol) of dezocine was weighed and placed in a 50ml single-necked round bottom flask, 3ml of tetrahydrofuran was added for dissolution, at this time, a colorless transparent solution was obtained, and after the reaction solution was placed in an ice-water bath for cooling, 227. Mu.L (1.633 mmol) of triethylamine was added, and then 206. Mu.L (0.898 mmol) of di-tert-butyl dicarbonate was dissolved in 2ml of tetrahydrofuran to obtain a solution, which was added dropwise to the reaction solution, and the temperature was slowly raised to 25℃and the reaction was continued with stirring for 4 hours. After the completion of the reaction, the reaction mixture was evaporated to dryness under reduced pressure to give 298mg of a colorless transparent solution, which was directly subjected to the next reaction.
2. 298mg (calculated as theory) of the product from the previous step (0.816 mmol) was dissolved in 15ml of dichloromethane, then 341 μl (2.448 mmol) of triethylamine, 163mg (1.630 mmol) of succinic anhydride and 30mg of DMAP were added, the reaction was stirred at room temperature for 16 hours, after the completion of the reaction the solvent was evaporated under reduced pressure, 15ml of purified water and 15ml of dichloromethane were added for extraction, the organic phase was collected, the aqueous phase was further extracted with 15ml of dichloromethane of x 2, the organic phases were combined, washed with 30ml of purified water and 30ml of saturated brine respectively, the organic phase was collected, dried, filtered and evaporated under reduced pressure to give a pale yellow oil which was separated by thin layer chromatography (solvent and eluent are absolute ethanol, developer is dichloromethane: 95% ethanol: 1, 4-dioxane: ammonia=10:8:1, product rf=0.4) to give another pale yellow oil 188mg (0.423 mmol).
3. 188mg (0.423 mmol) of the pale yellow oily substance from the previous step was dissolved in 3ml of trifluoroacetic acid and 3ml of dichloromethane, and the mixture was stirred at room temperature for 5 hours, and evaporated to dryness under reduced pressure to give 132mg of a yellow oily residue, which was directly taken to the next step.
4. Dissolving 152mg (calculated according to theoretical value) of yellow oily residue (0.423 mmol) in 5ml of benzene, placing in an ice-water bath for stirring, slowly adding 118 mu L (0.846 mmol) of trifluoroacetic anhydride, generating white smoke, then transferring to room temperature for stirring reaction for 1 hour, and transferring to an oil bath at 80 ℃ for stirring reflux reaction for 3 hours; after the reaction, the solvent was evaporated under reduced pressure, and 143mg (0.324 mmol) of yellow oily dezocine artificial hapten was separated by thin layer chromatography (solvent and eluent are absolute ethanol, developing solvent is dichloromethane: 95% ethanol: 1, 4-dioxane: ammonia water=10:8:1:1, product rf=0.3).
As shown in figure 1, the purity of the purified dezocine artificial hapten I is more than 99 percent, meets the purity requirement and can be used for the next reaction.
As shown in FIG. 2, the molecular ion peak of dezocine artificial hapten I obtained in example 1 has a mass-to-charge ratio (m/z) of 441.44, which is consistent with the theoretical molecular weight, and the structural formula of dezocine artificial hapten I can be preliminarily determined as formula I.
Example 2: preparation of dezocine artificial antigen
1. 143mg (0.324 mmol) of the dezocine artificial hapten is dissolved in 7.15ml of tetrahydrofuran, 56mg (0.486 mmol) of NHS and 124mg (0.648 mmol) of EDCI are added, the mixture is stirred at room temperature and reacts for 24 hours in a dark place, the mixture is centrifuged at 10000rpm for 15min, the precipitate is removed, the supernatant is taken and dried, and the residue is dissolved in 2.86ml of dimethyl sulfoxide to obtain an activated dezocine hapten solution.
2. 175mg of BSA was dissolved in 35ml of PBS buffer at a concentration of 0.01M to prepare a 5mg/ml PBS solution.
3. Slowly adding the activated dezocine hapten solution into the BSA solution prepared in the previous step, rapidly stirring the BSA solution in the adding process, and then sealing and keeping out of the light, stirring and reacting for 4 hours at room temperature.
4. The reaction solution was transferred to a dialysis bag, which was dialyzed against 0.5% sodium carbonate solution at ph=12.00 for two days, and then into 0.01M PBS for 3 days, twice daily. Centrifuging the dialyzate to remove precipitate to obtain dezocine artificial antigen, and storing at-20deg.C.
Example 3: preparation of Dezocine Xin Shan clone antibody
Purifying the prepared dezocine artificial antigen, diluting to 2mg/ml by using a phosphate buffer solution, mixing with Freund's adjuvant 1:1, taking 0.3mg of the artificial antigen mixed solution for injection into the back subcutaneous and thigh muscle of a healthy rabbit each time, injecting once every 3-4 weeks, collecting immunized fresh rabbit blood after 7-9 days of injection each time, centrifuging at 12000rpm for 15min by using a high-speed centrifuge, detecting the binding titer of the antibody obtained in serum by using an ELISA method, and when the antibody titer reaches 1: and when the serum is above 200, obtaining the dezote Xin Duo clone antibody by purifying serum, then utilizing B cell fusion tumor cells activated by mouse immunity, screening by a monoclonal technology, and separating and purifying mouse ascites to obtain the dezote Xin Shan clone antibody.
Example 4: preparation of dezote Xin Jiaoti gold test paper
1. Diluting the prepared dezocine artificial antigen with phosphate buffer solution to a concentration of 0.1-2 mg/ml, adding sucrose to play a role in biological protection, and uniformly spraying the mixed solution on an NC film in a spraying amount of 1.0-1.5 ul/cm to obtain a detection line; the sheep anti-rabbit IgG and the sheep anti-mouse IgG in the raw materials of the quality control line are respectively 0.5-2.0 mg/ml and 0.1-0.5 mg/ml, and are uniformly mixed and sprayed on the NC film by a spraying amount of 1.0-1.5 ul/cm after adding sucrose, and the NC film is prepared after being placed in an oven at about 45 ℃ for 16-22 hours.
2. Mixing the colloidal gold solution with the regulated pH value with the dezote Xin Shan clone antibody solution, adding the BSA solution for blocking after reacting for 15-20 minutes, reacting for 15-20 minutes again, centrifuging for 30 minutes at 10000rpm, and diluting and precipitating with a buffer solution to obtain the labeled dezote Xin Shan clone antibody raw material.
3. Adding the marked dezote Xin Shan clone antibody raw material, rabbit IgG and mouse IgG into diluent, trehalose and sucrose, diluting to a proper concentration, uniformly spraying the mixture on a polyester film with a spraying amount of 1.0-1.5 ul/cm, and placing the mixture in an oven at about 37 ℃ for 12-22 hours to obtain the gold mark pad after spraying.
4. And sequentially lapping the filter paper, the NC film, the gold mark pad and the sample pad on the PVC sheet paperboard from top to bottom, wherein the filter paper and the gold mark pad are lapped on the NC film, and the sample pad is lapped on the gold mark pad.
Detection principle: dezocine, a common clinical analgesic, is often injected into the patient's body, about 80% of which is excreted as urine, with 1% of the prototype and the others being conjugates of gluconic acid glycoside. The colloidal gold product of the invention mainly detects dezocine and metabolites thereof in urine, and adopts a competition method to detect dezocine and hapten thereof in urine or other body fluids. If the urine to be detected contains dezocine, the dezocine in the urine is combined with the marked dezocine Xin Shan clone antibody, the marked dezocine Xin Shan clone antibody is combined with the dezocine in the urine, so that the dezocine is not combined with the dezocine Xin Kangyuan coated on the detection line, the detection line is wireless, and the marked rabbit IgG and mouse IgG are combined with the goat anti-rabbit IgG and the goat anti-mouse IgG of the quality control line, and the quality control line is wired. If the urine contains dezocine and metabolites thereof, the C line is wired and the T line is wireless; if the urine does not contain dezocine and its metabolites, both the C line and the T line are wired.
The using method of the product comprises the following steps: the product is well preserved in dry and cool places, three drops of the sample solution to be tested are vertically dripped on a sample pad by a dropper, and the sample pad is read after waiting for 5 minutes. If the C line is wired, the T line is negative; if the C line is wired, the T line is wireless, and the result is positive.
Test example: dezote Xin Jiaoti gold test paper performance test
1. Sensitivity: dezocine solutions with different concentrations (0 ng/ml, 500ng/ml, 1000ng/ml and 1500 ng/ml) are prepared to test the performance of the dezocine Xin Jiaoti gold test paper product prepared by the invention, positive values are indicated by "+" in detection results, negative values are indicated by "-" in detection results, the dezocine solution with each concentration is repeatedly detected for 3 times, and the dezocine solution with different concentrations is detected by the dezocine Xin Jiaoti gold test paper as follows:
TABLE 1 detection of dezocine solutions at different concentrations by dezocine Xin Jiaoti gold test paper
| Dezocine concentration (ng/ml) | Detection result (+ or-) |
| 0 | -，-，- |
| 500 | -，-，- |
| 1000 | +，+，+ |
| 1500 | +，+，+ |
2. Stability: storing prepared dezote Xin Jiaoti gold test paper at-2 ℃ and 37 ℃ for 3 days respectively, taking dezote Xin Jiaoti gold test paper placed for 3 days at normal temperature as a control, respectively detecting dezote solutions with concentrations of 0ng/ml, 500ng/ml and 1500ng/ml, repeatedly detecting three times by adopting grades to divide the detected result intensities into ten intensities of G1-G10, and taking an average value, wherein the detection line intensity results of the dezote Xin Jiaoti gold test paper stored under three temperature conditions are as follows:
TABLE 2 detection line intensity results of Dizodiac Xin Jiaoti gold test paper stored under different temperature conditions
3. Specificity: the 10ug/ml standard substance solution of common drugs such as morphine, ketamine, methamphetamine and other drugs is respectively used as the to-be-detected liquid to be detected by using the product of the invention, the positive in the detection result is represented by "+", the negative is represented by "-", the detection is repeated three times, and the detection result of the dezodiac Xin Jiaoti gold test paper is as follows:
table 3 detection results of Dizodiac Xin Jiaoti gold test paper on common drugs
| Cross material (10 ug/ml) | Detection result (+ or-) |
| Morphine | -，-，- |
| Ketamine | -，-，- |
| Methamphetamine | -，-，- |
| Ephedrine | -，-，- |
| Pseudoephedrine | -，-，- |
| Diazepam | -，-，- |
| Phenobarbital | -，-，- |
| Tramadol | -，-，- |
| Benzoyl Aikangning | -，-，- |
| Procaine | -，-，- |
| Meisharone | -，-，- |
| Gatifloxacin | -，-，- |
| Ranitidine | -，-，- |
4. Detection rate: collecting 200 clinical urine samples, respectively using the dezote Xin Jiaoti gold test paper and the LC/MS test prepared by the invention, dividing the number of positive samples detected by the test paper by the number of positive samples detected by the LC/MS test to obtain the positive detection rate of the test paper product, wherein the result and the detection rate of the clinical urine samples detected by the dezote Xin Jiaoti gold test paper are as follows:
TABLE 4 detection results of Dizodiac Xin Jiaoti gold test paper on clinical urine samples
| Detection method | Number of positive results | Number of negative results |
| Dizoi Xin Jiaoti gold test paper | 39 | 161 |
| LC/MS | 42 | 158 |
The positive detection rate of the dezote Xin Jiaoti gold test paper is as follows: 39/42 x 100% = 92.85%
The performance test of the dezocine Xin Jiaoti gold test paper can obtain that the minimum detection limit of the test paper prepared by the invention can reach 1000ng/ml, the sensitivity is high, the test results of other common drugs and medicines are not crossed, the test paper has good specificity, the positive detection rate of the test paper for clinical urine samples is up to 92.85 percent, and dezocine in urine of patients with dezocine abuse can be timely detected, so that the dezocine abuse condition can be found, and the test paper plays a key role in medicine supervision and abuse prevention.
Claims (10)
1. The dezocine hapten is characterized in that the structural formula is shown as a formula (1):
2. a process for the preparation of dezocine hapten according to claim 1, characterized in that it comprises the steps of:
step (1), dezocine is dissolved in tetrahydrofuran, triethylamine and di-tert-butyl dicarbonate are added, and the reaction is carried out for 3 to 5 hours to obtain a product A;
step (2), dissolving the product A in dichloromethane, adding triethylamine, succinic anhydride and 4-dimethylaminopyridine DMAP, reacting for 14-16 hours at room temperature, and separating by thin layer chromatography to obtain oily substance B;
step (3), dissolving the oily matter B in a mixed solution of trifluoroacetic acid and dichloromethane, stirring and reacting for 4-6 hours, and evaporating to dryness to obtain oily residue C;
and (4) dissolving oily residue C in benzene, adding trifluoroacetic anhydride for reaction for 1-2 hours at room temperature, then carrying out oil bath reflux reaction for a period of time, and separating by thin layer chromatography to obtain the dezocine artificial hapten.
3. The preparation method according to claim 2, wherein the molar ratio of dezocine, triethylamine and di-tert-butyl dicarbonate in step (1) is (0.5-2): (1.0-3.5): (0.5-2.5).
4. A process according to claim 2 or 3, wherein the molar ratio of product a, triethylamine and succinic anhydride in step (2) is (0.5-1.5): (1-4): (1-3).
5. The process according to claim 2, wherein the molar volume ratio of oily substance B, trifluoroacetic acid and dichloromethane in step (3) is 0.141mmol:1mL:1mL; in the step (4), the molar ratio of the oily residue C to the trifluoroacetic anhydride is (0.8-1.3): (1.7-2.4).
6. The dezocine artificial antigen is characterized in that the dezocine artificial antigen is obtained by coupling and purifying the dezocine hapten with bovine serum albumin according to the claim 1, and the structural formula is shown as the formula (2):
7. the method for preparing the dezocine artificial antigen according to claim 6, wherein the method comprises the following steps:
the dezocine artificial hapten, N-hydroxysuccinimide NHS and carbodiimide EDCI are mixed and then placed at room temperature for reaction for 20-24 hours, then added into bovine serum albumin BSA solution for stirring reaction for 3-5 hours, and dialyzed in PBS with the pH value of 10.00-13.00, the sodium carbonate solution with the mass percent concentration of 0.3-0.7% and the PBS with the concentration of 0.01-0.02M in sequence, and the dezocine artificial antigen is obtained after centrifugal precipitation.
8. The method of claim 7, wherein the molar ratio of dezocine artificial hapten, N-hydroxysuccinimide and carbodiimide is (0.8-1.3): (1.2-1.8): (1.7-2.1).
9. A dezocine clone Xin Shan antibody obtained by animal immunization with the dezocine artificial antigen of claim 4.
10. A dezote Xin Jiaoti gold test paper comprises a sample pad, a gold-labeled pad, a detection line and a quality control line, and is characterized in that the detection line is obtained by taking the dezote Xin Kangyuan as a raw material spot film according to claim 4.
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