CN103288723A - Sulfonamide hapten, and preparation method and application thereof - Google Patents
Sulfonamide hapten, and preparation method and application thereof Download PDFInfo
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- CN103288723A CN103288723A CN2012100532061A CN201210053206A CN103288723A CN 103288723 A CN103288723 A CN 103288723A CN 2012100532061 A CN2012100532061 A CN 2012100532061A CN 201210053206 A CN201210053206 A CN 201210053206A CN 103288723 A CN103288723 A CN 103288723A
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Abstract
The invention discloses a hapten, and a preparation method and application thereof, particularly a sulfonamide hapten. The invention also discloses a preparation method and application of the hapten. The kit quick detection product established on the basis of the sulfonamide hapten is convenient to use and low in detection cost; and the detection method is efficient, accurate and quick, can be used for simultaneously detecting mass samples, and is suitable for on-site supervision of sulfonamide residues in animal-derived samples and screening of mass samples.
Description
Technical field
The present invention relates to a kind of haptens and its preparation method and application, be specifically related to sulfa drugs haptens and its preparation method and application.
Technical background
(Sulfonamides is that a class has the sulfanilic amide structure, is used for the chemotherapeutic agent of prevention and treatment bacterial infection disease SAs) to sulfa drugs.Because having wide spectrum, stable, economic, easy-to-use and unite the Trimethoprim effect and can improve tens of times characteristics, the medicine of Chang Yiya treatment concentration comes prophylactic generation as fodder additives, improve the transformation efficiency of feed and promote growth of animal, wherein Chang Yong SAs just has tens kinds.But there is serious side effects in such medicine, and the medium-term and long-term existence of human body can cause many bacteriums to sulfonamides deposits yields resistance, and potential carinogenicity is arranged, and has therefore caused great attention both domestic and external.National regulations such as China and the U.S., European Union in the animal food maximum residue limit(MRL) (MRLs) of total sulfanilamide (SN) and single sulfanilamide (SN) be 0.1mg/kg, wherein the MRLs of sulfamethazine (SM2) is 0.025 μ g/ml.
At present, the detection method of sulfa drug residue mainly contains microbiology method, physico-chemical analysis method and immunization etc.The microbiology method is easy, expense is low, but during operational cost and susceptibility, specificity all relatively poor, on qualitative, quantitative, all have difficulties, can not satisfy the needs of modern residue detection, even some detected result does not also reach the requirement of maximum residue limit(MRL); The physics and chemistry method often is used for measuring the drug residue of animal tissues as the conclusive evidence method, as gas-chromatography (GC), high performance liquid chromatography (HPLC), gas-matter coupling (GC-MS), liquid-matter coupling (HPLC-MS) etc., these method sensitivities, accurate, high specificity, resolution are good, can measure multiple medicine simultaneously, but need the complex pretreatment of expensive instrument, sample, loaded down with trivial details cost time-consuming, that detect higher, can not execute-in-place, and need professional's operation, so limited its application; By comparison, enzyme-linked immunoassay method has the advantages such as screening detection that specificity is good, highly sensitive, easy and simple to handle, the detection cost is low, be suitable for the batch sample, and required equipment is few, easy simple to operate, with low cost, can satisfy China livestock and poultry cultivation family, slaughterhouse, food enterprise, government function supervision department etc. better and carry out testing.
Summary of the invention
The purpose of this invention is to provide a kind of sulfa drugs haptens and preparation method thereof.
Sulfa drugs haptens provided by the invention, molecular structural formula is:
The haptenic preparation method of sulfa drugs provided by the invention comprises the steps:
A) in 25ml single port bottle, add 6-amino-nicotinic acid, ethanol and 12mol/L hydrochloric acid, reflux, reaction 8h, the TLC monitoring, reaction finishes, and removal of solvent under reduced pressure is dissolved in saturated NaHCO
3The aqueous solution, ethyl acetate extraction, anhydrous Na S
2O
4Drying, column chromatography (ethyl acetate/petroleum ether, 2/1, v/v) purifying;
B) in 25ml single port bottle, add a) products therefrom, triethylamine (Et
3N) and CH
2Cl
2, after the stirring and dissolving, add catalyzer 4-Dimethylamino pyridine (DMAP).Slowly splash into the dichloromethane solution of 4-acetylsulphanilyl chloride, the TLC monitoring, 5h reacts the aftertreatment that finishes, drying, column chromatography purification (ethyl acetate/petroleum ether, 1/10, v/v);
C) in 25ml single port bottle, add b) products therefrom and the 2mol/LNaOH aqueous solution, reflux, the TLC monitoring, reaction 10h finishes, and regulates pH4～5, has solid to separate out, and is the sulfa drugs haptens.
Another object of the present invention is the application of above-mentioned sulfa drugs haptens in immunodetection, specifically comprise the sulfa drugs antigen that is obtained by described sulfa drugs haptens and carrier protein couplet, reach the sulfa drugs antibody that is prepared by gained sulfa drugs antigen-immunized animal.
Wherein said carrier proteins can be mouse serum protein, thyroprotein, bovine serum albumin, rabbit anteserum albumen, human serum protein, ovalbumin, hemocyanin or Fibrinogen,
Described antibody is the sulfa drugs monoclonal antibody.
The present invention also provides the enzyme linked immunological kit by above-mentioned sulfa drugs Antibody Preparation, and the application of sulfa drug residue in detecting animal derived sample.
Sulfa drugs haptens provided by the invention both farthest kept the chemical structure of sulfanilamide (SN) parent nucleus, again by the chemosynthesis transformation introduced can with the protein coupling-COOH, synthetic method is simple, purity, productive rate are higher; As raw material, preparation is suitable for the antigen system immune animal of animal immune with this haptens, and the tiring of gained antibody, specificity, avidity are all relatively good; The antibody of gained is used for enzyme linked immunological kit; can detect Sulphadiazine Sodium, sulfamonomethoxine, sulfamethoxazole, Sulphathiazole, phthaloyl sulphur thiazole, ayerlucil, seven kinds of medicines of sulfapyridine simultaneously; easy to use, detect that cost is low, detection method efficient, accurately, fast, can detect large batch of sample simultaneously, be suitable for the on-site supervision of sulfa drug residue in the animal derived sample and the examination of great amount of samples.Sulfa drugs haptens of the present invention plays a significant role in the detection of sulfa drugs.
Description of drawings
Fig. 1: sulfa drugs haptens synthetic route chart
Fig. 2: sulfa drugs haptens hydrogen nuclear magnetic resonance spectrogram
Fig. 3: sulfa drugs enzyme linked immunological kit typical curve
Embodiment
Further set forth the present invention below in conjunction with specific embodiment.Should be understood that these embodiment only are used for explanation the present invention, and be not used for limiting the scope of the invention.
The haptenic synthetic and evaluation of embodiment 1 sulfa drugs
One, sulfa drugs haptenic synthetic (synthetic route such as Fig. 1)
A) in 25ml single port bottle, add 6-amino-nicotinic acid 0.87g, ethanol 20ml, 12mol/L hydrochloric acid 3.7ml, reflux, reaction 8h, the TLC monitoring, reaction finishes, and removal of solvent under reduced pressure is dissolved in saturated NaHCO
3The aqueous solution, ethyl acetate extraction, anhydrous Na S
2O
4Drying, column chromatography (ethyl acetate/petroleum ether, 2/1, v/v) purifying;
B) in 25ml single port bottle, add a) products therefrom 0.89g, triethylamine (Et
3N) 1.6ml, CH
2Cl
210ml after the stirring and dissolving, adds catalyzer 4-Dimethylamino pyridine (DMAP).Slowly splash into the 2ml dichloromethane solution of 4-acetylsulphanilyl chloride, the TLC monitoring, 5h reacts the aftertreatment that finishes, drying, column chromatography purification (ethyl acetate/petroleum ether, 1/10, v/v);
C) in 25ml single port bottle, add b) products therefrom 1g, 2mol/LNaOH aqueous solution 120ml, reflux, the TLC monitoring, reaction 10h finishes, and regulates pH4～5, has solid to separate out, and is the sulfa drugs haptens.
Two, the haptenic evaluation of sulfa drugs
Get synthetic sulfa drugs haptens through nuclear magnetic resonance hydrogen spectruming determining, as shown in Figure 2, three peaks in the collection of illustrative plates about 6.7～8.2ppm are the H fignal center on the pyridine ring, unimodal about 11ppm is the N--H peak on the sulfanilamide (SN) key, 13.1ppm about unimodal be the carboxyl peak, illustrate that haptens synthesizes successfully.
Sulfa drugs haptens and carrier proteins are carried out coupling obtain sulfa drugs antigen.
One, immunogenic preparation---sulfa drugs haptens-bovine serum albumin conjugate is synthetic
Get 12mg sulfa drugs haptens, be dissolved in 1ml N, in the dinethylformamide (DMF), obtain solution I; Get 15mg carbodiimide (EDC) and fully dissolve the back in adding I with 0.2ml water, stir 24h under the room temperature, obtain solution II; Take by weighing bovine serum albumin (BSA) 40mg, make it fully to be dissolved among the 3ml PBS (pH7.2), solution II dropwise slowly is added drop-wise in the protein solution, and under room temperature, stirs 24h; With 4 ℃ of dialysis of 0.01mol/L PBS 3 days, change dialyzate every day 3 times, to remove unreacted small-molecule substance, namely obtain the sulfa drugs immunogen; Packing, standby in-20 ℃ of preservations.
Two, the preparation of coating antigen---sulfa drugs haptens-ovalbumin conjugate is synthetic
Get 15mg sulfa drugs haptens with 0.8ml DMF dissolving, be cooled to 10 ℃, obtain solution I; Get isobutyl chlorocarbonate 10 μ l and add among the I 10 ℃ of stirring reaction 30min; Get 30mg ovalbumin 2.2ml 50mmol/LNa
2CO
3Dissolving, 10 ℃ of reaction 4h, 4 ℃ are spent the night then; With 4 ℃ of dialysis of 0.01mol/LPBS 3 days, change dialyzate every day 3 times, to remove unreacted small-molecule substance, namely obtain the sulfa drugs coating antigen; Packing, standby in-20 ℃ of preservations.
Three, the evaluation of sulfa drugs antigen
Carrier proteins, sulfa drugs haptens, sulfa drugs hapten-carrier protein conjugate are made into the solution of 0.5mg/ml with the PBS of pH7.4, return to zero with 0.01mol/L pH7.4PBS, with ultraviolet spectrophotometer in the interscan of wavelength 200～800nm scope, obtain the absorption curve of carrier proteins, sulfa drugs haptens, sulfa drugs hapten-carrier protein conjugate, and calculate its in conjunction with than.Found that different absorption curves appears in the three, show the success of sulfa drugs haptens and carrier protein couplet, the combination of haptens and bovine serum albumin is than being (18～22): 1, with the combination of ovalbumin than being (16～21): 1.
One, sulfa drugs MONOCLONAL ANTIBODIES SPECIFIC FOR
Animal immune: immunogen is injected in the Balb/c mouse body, and immunizing dose is 150 μ g/, makes it produce polyclonal antibody serum.
Cytogamy and cloning: after the mice serum measurement result is higher, get its splenocyte, merge in 8: 1 ratios and SP2/0 myeloma cell, adopt indirect competitive ELISA to measure cell conditioned medium liquid, screen positive hole.Utilize limiting dilution assay that cloning is carried out in positive hole, up to the hybridoma cell strain that obtains secrete monoclonal antibody.
Cell cryopreservation and recovery: the monoclonal hybridoma strain of sulfa drugs is made 1 * 10 with frozen storing liquid
9The cell suspension of individual/ml is in the medium-term and long-term preservation of liquid nitrogen.Take out frozen pipe during recovery, put into 37 ℃ of water-bath middling speeds immediately and melt, behind the centrifugal removal frozen storing liquid, move in the culturing bottle and cultivate.
The production of monoclonal antibody and purifying: the Balb/c mouse peritoneal is only injected sterilization paraffin oil 0.5ml/, the monoclonal hybridoma strain 6 * 10 of 7 days pneumoretroperitoneum injection sulfa drugss
5Individual/as only, to gather ascites after 7 days.Carry out the ascites purifying with sad-saturated ammonium sulphate method ,-20 ℃ of preservations.
Two, the monoclonal antibody mensuration of tiring
Measuring tiring of antibody with the indirect competitive ELISA method is 1: (50000～100000).
Indirect competitive ELISA method: with sulfa drugs haptens-ovalbumin conjugate coated elisa plate, add Sulphadiazine Sodium standard solution, ELIAS secondary antibody and monoclonal antibody working fluid, 25 ℃ of reaction 30min pour out liquid in the hole, with PBST washings washing 3～5 times, pat dry with thieving paper; Add substrate colour developing liquid, behind 25 ℃ of reaction 15min, add the stop buffer termination reaction; Set microplate reader and measure every hole absorbance in wavelength 450nm place.
Three, the specificity of monoclonal antibody
The ability that antibodies specific refers to its homospecificity antigen combination and comparison with such antigen-analogues ability, cross reacting rate commonly used is as judgement criteria.Cross reaction is more little, and the specificity of antibody is then more high.
This experiment is done serial dilution with Sulphadiazine Sodium, sulfamonomethoxine, sulfamethoxazole, Sulphathiazole, phthaloyl sulphur thiazole, ayerlucil, sulfapyridine, sulfamethazine, sulfadimethoxine, sulfaquinoxaline, 11 kinds of sulfa drugss of Sulphamerazine; carry out indirect competitive ELISA with monoclonal antibody respectively; the production standard curve is analyzed and is obtained IC
50, be calculated as follows cross reacting rate then:
The result shows that the cross reacting rate of each sulfa drugs is: Sulphadiazine Sodium 100.0%, sulfamonomethoxine 97.6%, sulfamethoxazole 169.0%, Sulphathiazole 188.0%, phthaloyl sulphur thiazole 62.8%, ayerlucil 103%, sulfapyridine 51.9%, sulfamethazine＜1%, sulfadimethoxine＜1%, sulfaquinoxaline＜1%, Sulphamerazine＜1%.Antibody of the present invention can detect Sulphadiazine Sodium, sulfamonomethoxine, sulfamethoxazole, Sulphathiazole, phthaloyl sulphur thiazole, ayerlucil, 7 kinds of sulfa drugss of sulfapyridine simultaneously.
One, the composition of enzyme linked immunological kit
(1) bag is by the enzyme plate of sulfa drugs hapten-carrier protein conjugate;
(2) the sheep anti mouse anti-antibody of usefulness horseradish peroxidase-labeled;
(3) sulfa drugs monoclonal antibody working fluid;
(4) the Sulphadiazine Sodium standard solution is 6 bottles, and concentration is respectively 0 μ g/L, 0.5 μ g/L, 1.5 μ g/L, 4.5 μ g/L, 13.5 μ g/L, 40.5 μ g/L;
(5) substrate colour developing liquid is made up of A liquid and B liquid, and A liquid is urea peroxide, and B liquid is tetramethyl benzidine;
(6) stop buffer is 2mol/L sulfuric acid;
(7) concentrated cleaning solution is pH7.2, contains the carbonate buffer solution of 0.8%～1.2% tween 20,0.01 ‰～0.03 ‰ Thiomersalate sanitas, 0.1～0.3mol/L, and described per-cent is percent weight in volume;
(8) concentrating redissolution liquid is pH7.6, contains the phosphate buffered saline buffer of 8%～12% casein, 0.1～0.3mol/L, and described per-cent is percent weight in volume.
The main agents of this test kit provides with the form of working fluid, and the method for inspection is convenient and easy, has specificity height, highly sensitive, characteristics such as tolerance range is high, accuracy height, adopts single stage method simultaneously, has saved detection time.
Two, enzyme linked immunological kit detects the application of actual sample
1. sample pre-treatments
(1) fresh milk pre-treating process
Get 50 μ l fresh milk samples, add in the 450 μ l redissolution working fluid mixing; Getting 50 μ l is used for analyzing
(2) tissue (chicken, pork, fish, shrimp) pre-treating process
Behind homogenizer homogeneous structure sample, take by weighing the equal pledge of 2.0 ± 0.05g to 50ml polystyrene centrifuge tube, the phosphate buffered saline buffer whirling motion that adds 4mlpH=6 becomes pasty state, adds the 6ml ethyl acetate again, vibration 1min, the centrifugal 5min of 4000rpm room temperature (20-25 ℃/68-77); Get the 3ml supernatant liquor to the clean glass test tube of 10ml, under 50～60 ℃ of water-bath nitrogen gas stream, dry up; Add the 1ml normal hexane, with vortex instrument whirling motion 30s, add 1ml again and redissolve working fluid, whirling motion 10s, the centrifugal 5min of 4000rpm room temperature (20-25 ℃/68-77); Remove upper strata normal hexane phase, take off layer water 50 μ l and be used for analyzing.
(3) honey pre-treating process
Take by weighing 1.0 ± 0.05g honey sample to 50ml polystyrene centrifuge tube; Add 1ml 1mol/L hydrochloric acid soln, vibrate to honey with vibrator and all dissolve; Put into 37 ℃ of incubators and hatch 30min; Take out, add the phosphate buffered saline buffer of 1ml 1mol/L sodium hydroxide solution and 1ml pH=6, add 2ml acetonitrile and 6ml ethyl acetate more respectively, with the vibrator 1min that vibrates, the centrifugal 10min of 4000rpm room temperature (20-25 ℃/68-77); Get the 4ml upper organic phase to the clean glass test tube of 10ml, under 50～60 ℃ of water-bath nitrogen gas stream, dry up; Add the dissolving of 0.5ml redissolution working fluid; Getting 50 μ l is used for analyzing.
(4) urine pre-treating process
Get 50 μ l urine specimens, add in the 150 μ l washing working fluid mixing; Getting 50 μ l is used for analyzing.
2. detect with test kit
In the enzyme plate micropore that is coated with the sulfa drugs envelope antigen, add Sulphadiazine Sodium standard solution or sample solution 50 μ l, the sheep anti mouse anti-antibody 50 μ l/ holes that add horseradish peroxidase-labeled immediately, add sulfa drugs monoclonal antibody working fluid 50 μ l/ holes again, behind cover plate membrane cover plate, put lucifuge reaction 30min in 25 ℃ of thermostat containers, pour out liquid in the hole, every hole adds 250 μ l washingss, pour out liquid in the hole behind the 30s, so repetitive operation is washed plate 5 times, pats dry with thieving paper; Every hole adds substrate colour developing liquid A liquid urea peroxide 50 μ l, substrate colour developing liquid B liquid tetramethyl benzidine (TMB) 50 μ l, mixing gently vibrates, with lucifuge colour developing 15min in the rearmounted 25 ℃ of thermostat containers of cover plate membrane cover plate, every hole adds stop buffer 2mol/L sulfuric acid 50 μ l, the mixing that vibrates gently at the 450nm place, is measured every hole absorbance (OD value) with the microplate reader wavelength set.
3. detected result analysis
With the absorbancy mean value (B) of the standard solution of each concentration that the obtains absorbance (B divided by first standard solution (0 standard)
0) multiply by 100% again, obtain the percentage absorbance.Logarithmic value with Sulphadiazine Sodium standard substance concentration (μ g/L) is X-axis, and the percentage absorbance is Y-axis, the drawing standard graphic representation, as shown in Figure 3.With the percentage absorbance of same way calculation sample solution, read the corresponding concentration of sample from typical curve, multiply by its corresponding extension rate, multiply by the cross reacting rate of every kind of medicine correspondence again, be the actual concentrations of sulfa drugs in the sample.
Three, the enzyme linked immunological kit technical parameter determines
Lowest detectable limit: 20 parts of dummies are detected, find concentration corresponding to each percentage absorbance from typical curve, add that with the mean value of 20 this sulfonamides of increment substrate concentrations 3 times of standard deviations represent detectability, the result must this method be limited to 5 μ g/L to the lowest detection of milk sample, lowest detection to animal tissues, honey sample is limited to 0.5 μ g/kg, and the lowest detection of urine specimen is limited to 2 μ g/L.
The accuracy that accuracy and precision: ELISA measures represents that with the rate of recovery precision is represented with the variation coefficient.Get dummy, with the sulfa drugs of three concentration of 50,100,200 μ g/kg it being added recovery test (adds concentration and exceeds the curved measurement scope, need take the circumstances into consideration sample to detect again after the dilution,), the result must this method is 85% ± 15% to the rate of recovery of milk sample, the rate of recovery to animal tissues's sample is 75% ± 10%, the rate of recovery to the honey sample is 90% ± 10%, the rate of recovery to urine specimen is 95% ± 15%, variation within batch coefficient＜15%, interassay coefficient of variation＜25%.
Test kit of the present invention can be preserved 12 months at least at 2～8 ℃ after measured.
Claims (9)
1. sulfa drugs haptens is characterized in that molecular structural formula is:
2. one kind prepares the haptenic method of the described sulfa drugs of claim 1, it is characterized in that comprising the steps:
A) in 25ml single port bottle, add 6-amino-nicotinic acid, ethanol and 12mol/L hydrochloric acid, reflux, reaction 8h, the TLC monitoring, reaction finishes, and removal of solvent under reduced pressure is dissolved in saturated NaHCO
3The aqueous solution, ethyl acetate extraction, anhydrous Na S
2O
4Drying, column chromatography (ethyl acetate/petroleum ether, 2/1, v/v) purifying;
B) in 25ml single port bottle, add a) products therefrom, triethylamine (Et
3N) and CH
2Cl
2, after the stirring and dissolving, add catalyzer 4-Dimethylamino pyridine (DMAP).Slowly splash into the dichloromethane solution of 4-acetylsulphanilyl chloride, the TLC monitoring, 5h reacts the aftertreatment that finishes, drying, column chromatography purification (ethyl acetate/petroleum ether, 1/10, v/v);
C) in 25ml single port bottle, add b) products therefrom and the 2mol/LNaOH aqueous solution, reflux, the TLC monitoring, reaction 10h finishes, and regulates pH4～5, has solid to separate out, and is the sulfa drugs haptens.
3. sulfa drugs antigen, it is to be obtained by the described sulfa drugs haptens of claim 1 and carrier protein couplet, it is characterized in that molecular structural formula is:
4. sulfa drugs antigen as claimed in claim 3, its preparation method comprises the steps:
Get the sulfa drugs haptens, be dissolved in N, in the dinethylformamide (DMF), obtain solution I; Get carbodiimide (EDC) water and fully dissolve among the back adding I, stir 24h under the room temperature, obtain solution II; Take by weighing carrier proteins, make it fully to be dissolved among the PBS (pH7.2), solution II dropwise slowly is added drop-wise in the protein solution, and under room temperature, stirs 24h; With 4 ℃ of dialysis of 0.01mol/L PBS 3 days, change dialyzate every day 3 times, namely obtain the sulfa drugs immunogen.
Get the sulfamido haptens and dissolve with DMF, be cooled to 10 ℃, obtain solution I; Get isobutyl chlorocarbonate and add among the I 10 ℃ of stirring reaction 30min; Get carrier proteins 50mmol/L Na
2CO
3Dissolving, 10 ℃ of reaction 4h, 4 ℃ are spent the night then; With 4 ℃ of dialysis of 0.01mol/L PBS 3 days, change dialyzate every day 3 times, namely obtain the sulfa drugs coating antigen.
5. sulfa drugs antibody, it is to be prepared by sulfa drugs immunogen immune animal.
6. antibody as claimed in claim 5 is characterized in that described antibody is the sulfa drugs monoclonal antibody.
7. sulfa drugs enzyme linked immunological kit by each described Antibody Preparation of claim 5-6.
8. sulfa drugs enzyme linked immunological kit as claimed in claim 7 is characterized in that sulfa drugs is Sulphadiazine Sodium, sulfamonomethoxine, sulfamethoxazole, Sulphathiazole, phthaloyl sulphur thiazole, ayerlucil, sulfapyridine.
9. a sulfa drugs enzyme linked immunological kit detects the application of sulfa drug residue in the animal derived sample.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109021094A (en) * | 2018-08-01 | 2018-12-18 | 陕西医药控股医药研究院有限公司 | A kind of method and test strips quickly detecting influenza Susceptible population using sialic acid artificial antigen |
| CN109824604A (en) * | 2019-02-25 | 2019-05-31 | 中国农业大学 | Sulfonamide hapten and artificial antigen and the preparation method and application thereof |
| CN110330488A (en) * | 2019-06-20 | 2019-10-15 | 深圳市易瑞生物技术股份有限公司 | A kind of piroxicam haptens and its preparation method and application |
| CN112830893A (en) * | 2019-10-28 | 2021-05-25 | 成都倍特药业股份有限公司 | ROR gamma inhibitor, preparation method and medical application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018454A (en) * | 2011-09-21 | 2013-04-03 | 北京勤邦生物技术有限公司 | Sulfanilamide drug chemiluminescence enzyme-linked immunodetection kit |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100344971C (en) * | 2005-11-03 | 2007-10-24 | 北京望尔生物技术有限公司 | ELISA kit for detecting sulfanilamides residue in animal derived food |
-
2012
- 2012-03-02 CN CN201210053206.1A patent/CN103288723B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018454A (en) * | 2011-09-21 | 2013-04-03 | 北京勤邦生物技术有限公司 | Sulfanilamide drug chemiluminescence enzyme-linked immunodetection kit |
Non-Patent Citations (3)
| Title |
|---|
| NURIA PASTOR-NAVARRO ET AL: "Development of a group-specific immunoassay for sulfonamides Application to bee honey analysis", 《TALANTA》, vol. 71, 7 July 2007 (2007-07-07), pages 923 - 933 * |
| 房超: "磺胺类药物残留酶免疫分析的研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》, no. 4, 15 December 2004 (2004-12-15) * |
| 段振娟 等: "磺胺母核结构半抗原的合成与表征", 《天津科技大学学报》, vol. 22, no. 2, 30 June 2007 (2007-06-30), pages 24 - 28 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109021094A (en) * | 2018-08-01 | 2018-12-18 | 陕西医药控股医药研究院有限公司 | A kind of method and test strips quickly detecting influenza Susceptible population using sialic acid artificial antigen |
| CN109824604A (en) * | 2019-02-25 | 2019-05-31 | 中国农业大学 | Sulfonamide hapten and artificial antigen and the preparation method and application thereof |
| CN109824604B (en) * | 2019-02-25 | 2020-06-23 | 中国农业大学 | Sulfonamide hapten and artificial antigen as well as preparation method and application thereof |
| CN110330488A (en) * | 2019-06-20 | 2019-10-15 | 深圳市易瑞生物技术股份有限公司 | A kind of piroxicam haptens and its preparation method and application |
| CN112830893A (en) * | 2019-10-28 | 2021-05-25 | 成都倍特药业股份有限公司 | ROR gamma inhibitor, preparation method and medical application thereof |
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| CN103288723B (en) | 2017-06-23 |
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