CN109232238A - 吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 - Google Patents
吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 Download PDFInfo
- Publication number
- CN109232238A CN109232238A CN201810991947.1A CN201810991947A CN109232238A CN 109232238 A CN109232238 A CN 109232238A CN 201810991947 A CN201810991947 A CN 201810991947A CN 109232238 A CN109232238 A CN 109232238A
- Authority
- CN
- China
- Prior art keywords
- gemfibrozil capsules
- haptens
- gemfibrozil
- capsules
- antibody
- 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.)
- Granted
Links
- 229960003627 gemfibrozil Drugs 0.000 title claims abstract description 103
- HEMJJKBWTPKOJG-UHFFFAOYSA-N Gemfibrozil Chemical compound CC1=CC=C(C)C(OCCCC(C)(C)C(O)=O)=C1 HEMJJKBWTPKOJG-UHFFFAOYSA-N 0.000 title claims abstract description 102
- 239000002775 capsule Substances 0.000 title claims abstract description 102
- 239000000427 antigen Substances 0.000 title claims abstract description 46
- 102000036639 antigens Human genes 0.000 title claims abstract description 46
- 108091007433 antigens Proteins 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000007788 liquid Substances 0.000 claims description 32
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 18
- 238000001514 detection method Methods 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004458 analytical method Methods 0.000 claims description 8
- 102000014914 Carrier Proteins Human genes 0.000 claims description 7
- 108010078791 Carrier Proteins Proteins 0.000 claims description 7
- 241001465754 Metazoa Species 0.000 claims description 7
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 229940098773 bovine serum albumin Drugs 0.000 claims description 6
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 6
- JODRRPJMQDFCBJ-UHFFFAOYSA-N 2-Hydroxy-4-methylbenzaldehyde Chemical compound CC1=CC=C(C=O)C(O)=C1 JODRRPJMQDFCBJ-UHFFFAOYSA-N 0.000 claims description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- 238000005352 clarification Methods 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 230000028993 immune response Effects 0.000 claims description 3
- 239000012074 organic phase Substances 0.000 claims description 3
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 229960001866 silicon dioxide Drugs 0.000 claims description 3
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 238000003756 stirring Methods 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 12
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000000034 method Methods 0.000 description 25
- 239000000523 sample Substances 0.000 description 21
- 238000002965 ELISA Methods 0.000 description 17
- 230000015572 biosynthetic process Effects 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 238000003786 synthesis reaction Methods 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 238000002835 absorbance Methods 0.000 description 8
- 239000011248 coating agent Substances 0.000 description 8
- 238000000576 coating method Methods 0.000 description 8
- 229940092253 ovalbumin Drugs 0.000 description 7
- 241001494479 Pecora Species 0.000 description 6
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 6
- 238000011084 recovery Methods 0.000 description 6
- 239000007790 solid phase Substances 0.000 description 6
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 239000012224 working solution Substances 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 108010058846 Ovalbumin Proteins 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 238000005138 cryopreservation Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 230000001900 immune effect Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- KZAMQAAMEOXHBU-UHFFFAOYSA-N 5-bromo-2,2-dimethylpentanoic acid Chemical compound OC(=O)C(C)(C)CCCBr KZAMQAAMEOXHBU-UHFFFAOYSA-N 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000005875 antibody response Effects 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 239000003547 immunosorbent Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- OJRHUICOVVSGSY-RXMQYKEDSA-N (2s)-2-chloro-3-methylbutan-1-ol Chemical compound CC(C)[C@H](Cl)CO OJRHUICOVVSGSY-RXMQYKEDSA-N 0.000 description 1
- ZGGHKIMDNBDHJB-NRFPMOEYSA-M (3R,5S)-fluvastatin sodium Chemical compound [Na+].C12=CC=CC=C2N(C(C)C)C(\C=C\[C@@H](O)C[C@@H](O)CC([O-])=O)=C1C1=CC=C(F)C=C1 ZGGHKIMDNBDHJB-NRFPMOEYSA-M 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- SUAUILGSCPYJCS-UHFFFAOYSA-N Musk ambrette Chemical compound COC1=C([N+]([O-])=O)C(C)=C([N+]([O-])=O)C=C1C(C)(C)C SUAUILGSCPYJCS-UHFFFAOYSA-N 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- RYMZZMVNJRMUDD-UHFFFAOYSA-N SJ000286063 Natural products C12C(OC(=O)C(C)(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 RYMZZMVNJRMUDD-UHFFFAOYSA-N 0.000 description 1
- AJLFOPYRIVGYMJ-UHFFFAOYSA-N SJ000287055 Natural products C12C(OC(=O)C(C)CC)CCC=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 AJLFOPYRIVGYMJ-UHFFFAOYSA-N 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000004411 aluminium Substances 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 238000003452 antibody preparation method Methods 0.000 description 1
- 229960001770 atorvastatin calcium Drugs 0.000 description 1
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical compound C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 description 1
- IIBYAHWJQTYFKB-UHFFFAOYSA-N bezafibrate Chemical compound C1=CC(OC(C)(C)C(O)=O)=CC=C1CCNC(=O)C1=CC=C(Cl)C=C1 IIBYAHWJQTYFKB-UHFFFAOYSA-N 0.000 description 1
- 229960000516 bezafibrate Drugs 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- KNHUKKLJHYUCFP-UHFFFAOYSA-N clofibrate Chemical compound CCOC(=O)C(C)(C)OC1=CC=C(Cl)C=C1 KNHUKKLJHYUCFP-UHFFFAOYSA-N 0.000 description 1
- 229960001214 clofibrate Drugs 0.000 description 1
- 239000003433 contraceptive agent Substances 0.000 description 1
- 230000002254 contraceptive effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 229960000868 fluvastatin sodium Drugs 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000118 hair dye Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 229940027941 immunoglobulin g Drugs 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 description 1
- 229960004844 lovastatin Drugs 0.000 description 1
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 description 1
- RLSSMJSEOOYNOY-UHFFFAOYSA-N m-cresol Chemical compound CC1=CC=CC(O)=C1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- AJLFOPYRIVGYMJ-INTXDZFKSA-N mevastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=CCC[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 AJLFOPYRIVGYMJ-INTXDZFKSA-N 0.000 description 1
- 229950009116 mevastatin Drugs 0.000 description 1
- BOZILQFLQYBIIY-UHFFFAOYSA-N mevastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CCC=C21 BOZILQFLQYBIIY-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920006389 polyphenyl polymer Polymers 0.000 description 1
- TUZYXOIXSAXUGO-PZAWKZKUSA-N pravastatin Chemical compound C1=C[C@H](C)[C@H](CC[C@@H](O)C[C@@H](O)CC(O)=O)[C@H]2[C@@H](OC(=O)[C@@H](C)CC)C[C@H](O)C=C21 TUZYXOIXSAXUGO-PZAWKZKUSA-N 0.000 description 1
- 229960001495 pravastatin sodium Drugs 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 229960002855 simvastatin Drugs 0.000 description 1
- RYMZZMVNJRMUDD-HGQWONQESA-N simvastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)C(C)(C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 RYMZZMVNJRMUDD-HGQWONQESA-N 0.000 description 1
- -1 small molecule compound Chemical class 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 125000003698 tetramethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- AQLJVWUFPCUVLO-UHFFFAOYSA-N urea hydrogen peroxide Chemical compound OO.NC(N)=O AQLJVWUFPCUVLO-UHFFFAOYSA-N 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C59/00—Compounds having carboxyl groups bound to acyclic carbon atoms and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
- C07C59/40—Unsaturated compounds
- C07C59/74—Unsaturated compounds containing —CHO groups
-
- 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
- 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
- C07K14/77—Ovalbumin
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Urology & Nephrology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Gastroenterology & Hepatology (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明提供了一种吉非罗齐半抗原和人工抗原,分子结构式分别为:本发明还公开了此半抗原、人工抗原和以此抗原制备的抗体的制备方法,以及半抗原、抗体的用途。本发明最终制成的抗体检测灵敏度高、特异性强。
Description
技术领域
本发明涉及选择一种具有—CHO的、又最大可能包含吉非罗齐原有结构的化合物作为吉非罗齐半抗原,以此半抗原制成的人工抗原、特异性抗体;以及此类半抗原、人工抗原、特异性抗体的用途,和半抗原、人工抗原、抗体的制备方法。
背景技术
药物及个人护理品(PPCPs)作为人类的必需品,种类繁杂,包括各类抗生素、人工合成麝香、止痛药、降压药、避孕药、催眠药、减肥药、发胶、染发剂和杀菌剂等。吉非罗齐(GEM)是一种血脂调节药物,用于治疗高血脂等疾病。GEM进入污水处理厂后不能被完全去除,随着出水可以进入到水环境中,造成二次污染。由于其在环境介质中的浓度水平较低(ng/L~μg/L),因此往往被人们忽视。虽然PPCPs的半衰期较短,但由于有源源不断的输入源头,导致其在环境介质中呈现“伪持续存在”的状态,长期暴露对生物和人类健康造成潜在危害。随着人们环境保护意识的增强,吉非罗齐等“新兴污染物”在近些年来越来越受到广泛的关注,而建立一系列速度快、成本低、准确性高的检测方法显得极为迫切。
目前对吉非罗齐的检测主要为仪器方法,如液相色谱法、液相色谱-质谱法等。仪器检测法存在样品前处理繁琐、检测时间长、仪器贵重等缺点,所以在我国无法得到广泛应用,并且不符合现场检测“在短时间内低成本对大量样品进行准确检测和筛选”的要求。酶联免疫吸附测定技术Enzyme-Linked Immunosorbent Assay(ELISA),是将免疫技术与现代测试手段相结合而建立的一种超微量的测定方法,具有成本低、速度快、灵敏度高、仪器设备简单的特点,适合大批量样品的快速分析。而免疫学检测方法的基础是相应的高特异性、高亲和力的抗体的制备,而吉非罗齐作为小分子化合物,本身不具备免疫原性,因此,其半抗原的结构改造和全抗原合成是建立免疫速测技术的关键和难点之一。
发明内容
针对现有技术中存在的不足之处,本发明提供一种能最大程度保留吉非罗齐的化学结构,又具有一定长度连接臂的半抗原以及这种半抗原的制备方法;以此半抗原制备的人工抗原、检测灵敏度高和特异性强的抗体;以及此半抗原、抗体的用途。
本发明为达到以上目的,是通过这样的技术方案来实现的:提供一种吉非罗齐半抗原,它的分子结构式为:
本发明的半抗原以2-羟基-4-甲基苯甲醛为起始原料,与5-溴-2,2-二甲基戊酸反应生成醛基吉非罗齐,即为吉非罗齐半抗原。反应式如下:
本发明的吉非罗齐半抗原的制备方法进一步描述如下:
取2-羟基-4-甲基苯甲醛0.5g,加乙腈50mL溶解,澄清,加碳酸钾0.61g,加5-溴-2,2-二甲基戊酸0.84g,油浴加热回流反应3h;停止反应,冷却至室温,旋蒸,除去乙腈,加水50mL,乙酸乙酯100mL×3,萃取三次,合并有机相,上硅胶柱,用体积比1:3的乙酸乙酯-石油醚层析纯化,即可得到吉非罗齐半抗原。
上述方法制得的吉非罗齐半抗原,具有吉非罗齐特征结构的同时又具有可以与蛋白质偶联的—CHO结构。
本发明还提供了一种依靠上述吉非罗齐半抗原制得的吉非罗齐人工抗原,它的分子结构式为:
所述载体蛋白为牛血清白蛋白。
本发明的吉非罗齐人工抗原的制备方法描述如下:
取吉非罗齐半抗原9.8mg,加乙醇0.2mL溶解,得到A液;取载体蛋白50mg,加0.1mol/L碳酸氢钠溶液3mL溶解,得到B液;将A液滴加到B液中,4℃搅拌24h,生理盐水透析纯化3d,每天换液3次,分装,-20℃保存备用。
本发明同时提供了上述吉非罗齐半抗原的用途,是用作动物免疫的抗原体系的原料。
本发明同时提供了上述吉非罗齐人工抗原免疫白鼠所得到的、能与吉非罗齐发生特异性免疫反应的吉非罗齐单克隆抗体,及其在检测吉非罗齐残留中的应用。
基于吉非罗齐—COOH连接臂可能是其活性基团,对维系半抗原的免疫特性和特征结构十分重要,或若—COOH直接与载体连接后半抗原的特征结构易受载体的局部微化学环境或空间位阻的干扰,影响机体免疫系统的识别,因此,以2-羟基-4-甲基苯甲为原料,经过与5-溴-2,2-二甲基戊酸反应,合成了带有醛基的半抗原,最大可能保留了原来吉非罗齐的分子结构,使吉非罗齐分子的化学结构与电子分布几乎不受影响,这为获得对吉非罗齐有高度特异性的抗体提供了保证。
本发明中合成的吉非罗齐半抗原,既最大程度地保留了吉非罗齐的化学结构,又具有一定长度的连接臂,用这一半抗原制备的免疫抗原去免疫动物,所得的抗体的效价、特异性、亲和力都比较好,所得的抗体用于ELISA方法检测吉非罗齐,灵敏度可达1μg/L,与其他调血脂类药物的交叉反应率低。
附图说明
图1吉非罗齐半抗原合成路线图
图2吉非罗齐ELISA竞争标准曲线图
具体实施方式
下面结合附图和具体实施例进一步详细说明本发明。下述实施例中所使用的实验方法如无特殊说明,均为常规方法;所使用的材料、试剂等,如无特殊说明,为可从商业途径得到的试剂和材料。
实施例1:半抗原合成
一种吉非罗齐半抗原,分子结构式为:
是用作动物免疫的抗原体系的原料。
上述吉非罗齐半抗原的制备方法如下(合成路线见附图1):
取2-羟基-4-甲基苯甲醛0.5g,加乙腈50mL溶解,澄清,加碳酸钾0.61g,加5-溴-2,2-二甲基戊酸0.84g,油浴加热回流反应3h;停止反应,冷却至室温,旋蒸,除去乙腈,加水50mL,乙酸乙酯100mL×3,萃取三次,合并有机相,上硅胶柱,用体积比1:3的乙酸乙酯-石油醚层析纯化,即可得到吉非罗齐半抗原。
实施例2:人工抗原合成和鉴定
由实施例1所述吉非罗齐半抗原制成的一种吉非罗齐人工抗原,分子结构式为:
1、免疫原的合成
免疫原的合成方法如下:
取吉非罗齐半抗原9.8mg,加乙醇0.2mL溶解,得到A液;取载体蛋白50mg,加0.1mol/L碳酸氢钠溶液3mL溶解,得到B液;将A液滴加到B液中,4℃搅拌24h,生理盐水透析纯化3d,每天换液3次,分装,-20℃保存备用。
人工抗原的鉴定:
按合成吉非罗齐免疫原反应所用半抗原、载体蛋白与偶联产物的比例,进行紫外(200~400nm)扫描测定,通过比较三者分别在260nm和280nm的吸光值计算其结合比。偶联物吉非罗齐半抗原-BSA的最大吸收峰与吉非罗齐半抗原、BSA的最大吸收峰相比发生了明显的变化,表明人工抗原吉非罗齐半抗原-BSA的合成是成功的。经计算,半抗原与BSA的结合比为13:1。
2、包被原的合成
包被原的合成方法同免疫原的合成,将9.8mg吉非罗齐半抗原调整为4.8mg,50mg牛血清白蛋白替换为50mg卵清蛋白(OVA)。
人工抗原的鉴定:
按合成吉非罗齐包被原反应所用半抗原、载体蛋白与偶联产物的比例,进行紫外(200~400nm)扫描测定,通过比较三者分别在260nm和280nm的吸光值计算其结合比。偶联物吉非罗齐半抗原-OVA的最大吸收峰与吉非罗齐半抗原、OVA的最大吸收峰相比发生了明显的变化,表明人工抗原吉非罗齐半抗原-OVA的合成是成功的。经计算,半抗原与OVA的结合比为10:1。
实施例3:单克隆抗体制备
一种吉非罗齐特异性抗体,是将实施例2所述吉非罗齐免疫原免疫白鼠所得到的、能与吉非罗齐发生特异性免疫反应的单克隆免疫球蛋白G,用于检测吉非罗齐残留。
上述吉非罗齐单克隆抗体的制备方法如下:
1)动物免疫:将免疫原注入到Balb/c小鼠体内,免疫剂量为150μg/只,使其产生抗血清。
2)细胞融合和克隆化:取产生特异性抗体的Balb/c小鼠脾细胞与骨髓瘤细胞SP20融合,采用间接竞争酶联免疫分析方法测定细胞上清液,筛选阳性孔。利用有限稀释法对阳性孔进行克隆化,得到并建立产单克隆抗体的杂交瘤细胞株。
3)细胞冻存和复苏:取处于对数生长期的杂交瘤细胞用冻存液制成细胞悬液,分装于冻存管,在液氮中长期保存。复苏时取出冻存管,立即放入37℃水浴中速融,离心去除冻存液后,移入培养瓶内培养。
4)单克隆抗体的制备与纯化:采用体内诱生法,将Balb/c小鼠(8周龄)腹腔注入灭菌石蜡油,7~14天后腹腔注射杂交瘤细胞,7~10天后采集腹水。经辛酸-饱和硫酸铵法进行腹水纯化,纯度经SDS-PAGE电泳鉴定,小瓶分装,-20℃保存。
抗体的特异性测定:
用具有多种抗原决定簇的免疫原(蛋白质或多肽)制备的抗体,其中含的抗体分子往往是混合体。当有甲、乙两种抗原,其分子结构中具有相同或部分相同的抗原决定簇时,甲抗原可与乙抗原的抗体反应,而乙抗原也可与甲抗原的抗体反应,称为交叉反应。抗体的特异性就是指它同特异性抗原结合的能力与同该抗原类似物结合能力的比较。常用交叉反应活性作为评价的重要标准。交叉反应越小,抗体的特异性则越好。
将特异性抗原及其类似物做系列稀释,分别与同一种吉非罗齐半抗原-BSA抗体,按制作标准曲线同样方法制作标准曲线,并在曲线上找出抑制率50%的剂量和类似物抑制率50%时的用量。然后计算出各类似物的交叉反应率。
抗吉非罗齐半抗原-BSA抗体对吉非罗齐及各类似物的交叉反应率:吉非罗齐为100%,普伐他汀钠、苯扎贝特、阿托伐他汀钙、氟伐他汀钠、氯贝丁酯、美伐他汀、洛伐他汀、辛伐他汀均<0.1%。由此可见,所制备的抗体特异性较好。
实施例4:吉非罗齐酶联免疫吸附测定方法的建立与应用
一、吉非罗齐ELISA测定方法的基本原理
采用间接竞争酶联免疫分析方法,即将吉非罗齐分子与大分子载体(如蛋白质)偶联制得的复合物作为包被原吸附于固相载体(96孔酶标板)上,制备成固相抗原,然后加入待测样品和相应抗体。固相抗原、待测样品中的吉非罗齐与抗体进行竞争结合反应,待测样品中吉非罗齐含量多,则被结合在固相抗原上的抗体少,反之结合在固相抗原上的抗体多,然后加入酶标二抗,最后用底物进行显色加以测定。当抗体量一定时,加入的待测样本中吉非罗齐量越多,与固相抗原结合的抗体就越少,与抗体结合的酶标二抗也越少,发色反应就减弱,反之,则发色反应增强,因而可根据已知吉非罗齐量的标准曲线和待测样品的吸光度值,推算出待测样品中吉非罗齐的浓度。
二、吉非罗齐ELISA测定方法中涉及的组分
(1)包被有吉非罗齐人工抗原(吉非罗齐半抗原-卵清蛋白偶联物)的酶标板;
(2)吉非罗齐系列标准品溶液:浓度分别为0μg/L、1μg/L、3μg/L、9μg/L、27μg/L、81μg/L;
(3)抗体工作液:实施例3中所述的吉非罗齐单克隆抗体工作液;
(4)酶标二抗:用辣根过氧化物酶标记的羊抗鼠抗抗体;
(5)底物显色液:由A液和B液组成,A液为2%过氧化脲的水溶液,B液为1%四甲基联苯胺的水溶液;
(6)终止液:2mol/L硫酸水溶液;
(7)洗涤液:含有1.0%吐温-20、0.02‰叠氮化钠,pH值为7.4、0.05mol/L的磷酸盐缓冲液;
(8)复溶液:含有5%牛血清白蛋白,pH值为7.4、0.02mol/L的磷酸盐缓冲液。
三、吉非罗齐ELISA测定方法中各组分的制备
1、包被有吉非罗齐人工抗原(吉非罗齐半抗原-卵清蛋白偶联物)的酶标板的制备
用包被缓冲液将实施例2中所述吉非罗齐包被原稀释至0.2μg/mL,包被96孔聚苯乙烯酶标板,每孔100μL,37℃温育2h,倾去包被液,用洗涤液洗涤3次,每次30s,拍干,然后在每孔加入200μL封闭液,37℃温育2h,倾去孔内液体,干燥后用铝膜真空密封保存。
所用的包被缓冲液和封闭液如下:
包被缓冲液:pH9.6、0.05mol/L的碳酸盐缓冲液;
封闭液:含有0.5%马血清、0.1%叠氮化钠、3%酪蛋白,pH值为7.2、0.02mol/L的磷酸盐缓冲液。
2、吉非罗齐系列标准品溶液的制备
用标准品稀释液将吉非罗齐标准品稀释至浓度分别为0μg/L、1μg/L、3μg/L、9μg/L、27μg/L、81μg/L;标准品稀释液为含有5%牛血清白蛋白,pH值为7.4、0.02mol/L的磷酸盐缓冲液。
3、抗体工作液的制备
用抗体稀释液将实施例3中所述的吉非罗齐单克隆抗体稀释1000倍,得到抗体工作液;抗体稀释液为含有2.5%酪蛋白和0.03‰叠氮化钠的磷酸盐缓冲液。
4、酶标二抗的制备
(1)羊抗鼠抗抗体的制备
以羊作为免疫动物,以鼠源抗体为免疫原对无病原体羊进行免疫,得到羊抗鼠抗抗体。
(2)酶标二抗的制备
将羊抗鼠抗抗体与辣根过氧化物酶采用过碘酸钠法进行偶联,然后用酶标二抗稀释液将其稀释500倍;所述酶标二抗稀释液为含0.5%牛血清白蛋白,pH值为7.4、0.02mol/L的磷酸盐缓冲液。
四、用上述吉非罗齐ELISA测定方法检测样品中残留的吉非罗齐
(一)样品前处理
用复溶液将水样进行2倍稀释。
(二)检测
向包被有吉非罗齐半抗原-卵清蛋白偶联物的酶标板微孔中加入系列标准品溶液或样品溶液50μL,随即加入酶标二抗50μL,再加入单克隆抗体工作液50μL,轻轻振荡混匀,用盖板膜盖板后置25℃避光环境中反应30min;倒出孔内液体,每孔加入洗涤液250μL,10s后倒出孔内液体,如此重复操作共洗板5次,用吸水纸拍干;每孔加入底物显色液A液和B液各50μL,轻轻振荡混匀,用盖板膜盖板后置25℃避光环境中反应15min;每孔加入终止液50μL,轻轻振荡混匀,设定酶标仪于450nm处,测定每孔的吸光度值。
(三)检测结果分析
用所获得的每个浓度的标准品溶液的吸光度平均值(B)除以第一个标准品溶液(0标准)的吸光度平均值(B0)再乘以100%,得到百分吸光度值。
以吉非罗齐标准品浓度的对数值为X轴,百分吸光度值为Y轴,绘制标准曲线图(图2)。用同样的方法计算样品溶液的百分吸光度值,相对应每一个样品的浓度,则可从标准曲线上读出样品中吉非罗齐的残留量。本发明中检测结果的分析也可以采用回归方程法,计算出样品溶液浓度。本发明中检测结果的分析还可以利用计算机专业软件,此法更便于大量样品的快速分析,整个检测过程只需1.0h可以完成。
五、上述吉非罗齐ELISA测定方法检测效果评价
(一)最低检测限
取不含吉非罗齐的阴性水样20份,将样品进行前处理后用上述吉非罗齐ELISA测定方法对其进行检测,以20份样品检测浓度的平均值加上3倍标准差表示检测限。
结果表明,该方法对水样的最低检测限为2μg/L。
(二)准确度和精密度
以回收率作为准确度评价指标,重复测定某一浓度样品的检测结果相对标准偏差(RSD%)作为精密度评价指标。计算公式为:回收率(%)=实际测定值/理论值×100%,其中理论值为样品的添加浓度;相对标准偏差RSD%=SD/X×100%,其中SD为标准偏差,X为测定数据的平均值。
向不含吉非罗齐的水样中分别添加吉非罗齐,使其终浓度为2μg/L、4μg/L、8μg/L,将添加后的样品进行前处理后用上述吉非罗齐ELISA测定方法对其进行检测,选用3批试剂,每个浓度做5个平行,分别计算回收率和批内、批间RSD,结果如表1所示。结果表明,水样的平均回收率在80%~100%,批内、批间RSD均在10%以内。
表1准确度和精密度实验结果
(三)保存期
上述吉非罗齐ELISA测定方法中所涉及到的主要试剂最终以工作液的形式提供,组装成试剂盒,大大降低了移液和操作的误差,同时体积小,易于携带和运送至终端客户处,更适合应用于现场操作检测和大批量样本检测,也节约了快递运输成本费用。
试剂盒保存条件为2-8℃,经过12个月的测定,试剂盒的最大吸光度值(零标准)、50%抑制浓度、吉非罗齐添加回收率均在正常范围内。考虑到运输和使用过程中会有非正常保存条件出现,将试剂盒在37℃下放置8天进行加速老化实验,结果该试剂盒的各项指标完全符合要求;考虑到试剂盒冷冻情况发生,将试剂盒在-20℃冰箱中放置8天,结果该试剂盒的各项指标也完全正常。从以上结果可得出,试剂盒可以在2-8℃至少保存12个月以上。
Claims (7)
1.一种吉非罗齐半抗原,其特征在于它的分子结构式为:
2.一种权利要求1所述吉非罗齐半抗原的制备方法,其特征在于包括下列步骤:
取2-羟基-4-甲基苯甲醛0.5g,加乙腈50mL溶解,澄清,加碳酸钾0.61g,加5-溴-2,2-二甲基戊酸0.84g,油浴加热回流反应3h;停止反应,冷却至室温,旋蒸,除去乙腈,加水50mL,乙酸乙酯100mL×3,萃取三次,合并有机相,上硅胶柱,用体积比1:3的乙酸乙酯-石油醚层析纯化,即可得到吉非罗齐半抗原。
3.一种吉非罗齐人工抗原,其特征在于它的分子结构式为:
所述载体蛋白为牛血清白蛋白。
4.一种权利要求3所述吉非罗齐人工抗原的制备方法,其特征在于包括下列步骤:
取吉非罗齐半抗原9.8mg,加乙醇0.2mL溶解,得到A液;取载体蛋白50mg,加0.1mol/L碳酸氢钠溶液3mL溶解,得到B液;将A液滴加到B液中,4℃搅拌24h,生理盐水透析纯化3d,每天换液3次,分装,-20℃保存备用。
5.一种权利要求1所述吉非罗齐半抗原在制备动物免疫的抗原体系方面的应用,其特征在于是用作动物免疫的抗原体系的原料。
6.一种采用权利要求3所述吉非罗齐人工抗原免疫白鼠所得到的、能与吉非罗齐发生特异性免疫反应的吉非罗齐单克隆抗体。
7.一种权利要求6所述吉非罗齐单克隆抗体在检测吉非罗齐残留中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810991947.1A CN109232238B (zh) | 2018-08-29 | 2018-08-29 | 吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810991947.1A CN109232238B (zh) | 2018-08-29 | 2018-08-29 | 吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN109232238A true CN109232238A (zh) | 2019-01-18 |
CN109232238B CN109232238B (zh) | 2022-05-17 |
Family
ID=65069360
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810991947.1A Active CN109232238B (zh) | 2018-08-29 | 2018-08-29 | 吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109232238B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109970550A (zh) * | 2019-03-26 | 2019-07-05 | 深圳市易瑞生物技术股份有限公司 | 山梨酸半抗原、人工抗原、抗体及其合成方法和应用 |
CN113563181A (zh) * | 2021-08-11 | 2021-10-29 | 浙江精进药业有限公司 | 吉非罗齐杂质的去除方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1326358A (zh) * | 1998-08-26 | 2001-12-12 | 葛兰素集团有限公司 | Dna接种的方法 |
-
2018
- 2018-08-29 CN CN201810991947.1A patent/CN109232238B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1326358A (zh) * | 1998-08-26 | 2001-12-12 | 葛兰素集团有限公司 | Dna接种的方法 |
Non-Patent Citations (4)
Title |
---|
M. CERMOLA ET AL.,: ""Phototransformation of fibrate drugs in aqueous media"", 《ENVIRON CHEM LETT》 * |
SEDA DEMIRELTOPEL ET AL.,: ""Synthesis and characterization of Bodipy functionalized magnetic iron oxide nanoparticles for potential bioimaging applications"", 《COLLOIDS AND SURFACES B: BIOINTERFACES》 * |
刘丽主编: "《胶体金免疫层析技术》", 30 September 2017, 河南科学技术出版社 * |
贾连群,雷萍主编: "《现代基础医学理论与技术进展》", 31 August 2015, 中国医药科技出版社 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109970550A (zh) * | 2019-03-26 | 2019-07-05 | 深圳市易瑞生物技术股份有限公司 | 山梨酸半抗原、人工抗原、抗体及其合成方法和应用 |
CN109970550B (zh) * | 2019-03-26 | 2021-09-07 | 深圳市易瑞生物技术股份有限公司 | 山梨酸半抗原、人工抗原、抗体及其合成方法和应用 |
CN113563181A (zh) * | 2021-08-11 | 2021-10-29 | 浙江精进药业有限公司 | 吉非罗齐杂质的去除方法 |
CN113563181B (zh) * | 2021-08-11 | 2024-04-26 | 浙江精进药业有限公司 | 吉非罗齐杂质的去除方法 |
Also Published As
Publication number | Publication date |
---|---|
CN109232238B (zh) | 2022-05-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101571539B (zh) | 检测头孢类药物的酶联免疫试剂盒及其应用 | |
CN102967709B (zh) | 检测玉米赤霉烯酮药物的酶联免疫试剂盒及其应用 | |
CN101354401B (zh) | 头孢氨苄残留酶联免疫检测试剂盒及其应用 | |
CN101256188A (zh) | 检测林可霉素药物的酶联免疫试剂盒及其应用 | |
CN106771137B (zh) | 检测尼卡巴嗪的酶联免疫试剂盒及其应用 | |
CN101776685B (zh) | 检测三甲氧苄胺嘧啶药物的酶联免疫试剂盒及其应用 | |
CN102080067B (zh) | 一种检测呕吐毒素的方法及其专用试剂盒 | |
CN109232238A (zh) | 吉非罗齐半抗原、人工抗原和抗体及其制备方法和用途 | |
CN109369435A (zh) | 托芬那酸半抗原、人工抗原和抗体及其制备方法和用途 | |
CN101907623B (zh) | 一种检测庆大霉素和/或小诺霉素的方法及其专用量子点荧光免疫试剂盒 | |
CN102367270A (zh) | 环孢霉素a半抗原的制备方法及环孢霉素a的酶联免疫定量检测试剂盒 | |
CN105785012A (zh) | 检测利巴韦林的酶联免疫试剂盒及其应用 | |
Zhang et al. | Rapid monitoring of dipropyl phthalate in food samples using a chemiluminescent enzyme immunoassay | |
CN101936985A (zh) | 一种检测己烯雌酚的方法及其专用化学发光免疫试剂盒 | |
CN103364553A (zh) | 检测硝基咪唑类药物的酶联免疫试剂盒及其应用 | |
CN103018451A (zh) | 检测氯霉素的酶联免疫试剂盒及其应用 | |
CN100492009C (zh) | 检测氟甲喹药物的酶联免疫试剂盒及方法 | |
CN106872681B (zh) | 丁胺卡那霉素和卡那霉素二合一快速检测酶联免疫试剂盒及其应用 | |
CN104569398B (zh) | 检测乙氧基喹啉的酶联免疫试剂盒及其应用 | |
CN101955537A (zh) | 一种莱克多巴胺抗体及其应用 | |
CN105158472A (zh) | 检测甲霜灵残留的酶联免疫试剂盒及使用方法 | |
CN109813880A (zh) | 一种检测三甲氧苄胺嘧啶残留的酶联免疫试剂盒 | |
CN109180507B (zh) | 酮洛芬半抗原、人工抗原和抗体及其制备方法和用途 | |
CN1308686C (zh) | 一种检测氯霉素的试剂盒 | |
CN103508910B (zh) | β-兴奋剂类半抗原及其制备方法和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |