CN104292335A - Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 - Google Patents
Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 Download PDFInfo
- Publication number
- CN104292335A CN104292335A CN201410497912.4A CN201410497912A CN104292335A CN 104292335 A CN104292335 A CN 104292335A CN 201410497912 A CN201410497912 A CN 201410497912A CN 104292335 A CN104292335 A CN 104292335A
- Authority
- CN
- China
- Prior art keywords
- thiabendazole
- cell strain
- hybridoma cell
- monoclonal antibody
- add
- 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
- 0 CCO*(C(N)=CSC=C)O Chemical compound CCO*(C(N)=CSC=C)O 0.000 description 6
- UOEZLTHJROMVTN-UHFFFAOYSA-N CCOC(C(CSC=C)N)=O Chemical compound CCOC(C(CSC=C)N)=O UOEZLTHJROMVTN-UHFFFAOYSA-N 0.000 description 1
- JSYBAZQQYCNZJE-UHFFFAOYSA-N Nc(cc1N)ccc1N Chemical compound Nc(cc1N)ccc1N JSYBAZQQYCNZJE-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a hybridoma cell strain FQ-B12, an anti-benzimidazole sterilizing agent namely a thiabendazole monoclonal antibody secreted by the cell strain, and an application of the hybridoma cell strain FQ-B12. The hybridoma cell strain FQ-B12 is collected in China Center for Type Culture Collection (CCTCC), and has the collection number of CCTCC NO. C201491. The monoclonal antibody secreted by the hybridoma cell strain FQ-B12 can be used for identifying thiabendazole, is high in sensitivity and good in specificity, and has the 50% inhibition concentration (IC50) of 6.0ng.mL<-1> to thiabendazole. An enzyme-linked immunosorbent assay method researched by using the monoclonal antibody can achieve the detection of thiabendazole in water, apples and pears.
Description
Technical field
The present invention relates to hybridoma cell strain FQ-B12, the monoclonal antibody of the anti-benzimidazoles sterilant thiabendazole of its secretion and application.
Background technology
Benzimidazoles agricultural chemicals is the important curative fungicide agent of a class, due to its drug effect significantly, toxicity is lower, be widely used in the Production of fruit of fungal infection by as broad spectrum systemic fungicide.According to " 2007 agricultural chemicals management information compilation ", derosal, F-1991 and thiabendazole are the main species of benzimidazoles agricultural chemicals.The major toxicity of this kind of agricultural chemicals is teratogenesis, is first proposed in 1973 by Lapras M etc.Its longevity of residure is longer, degrades comparatively slow, have certain inrichment in physical environment.And edible this kind of material is larger to the body harm of the mankind when reaching doses.
China is the first in the world Production of fruit big country, and fruit safety occupies very consequence in food safety.Along with the development of fruit industry, healthy to people of pesticide residues in fruits and the serious harm that exceeds standard.Diphenylimidazolidin-4-one series bactericidal agent, as thiabendazole, is widely used in the control of the various crop vegetative period such as fruit and shelf lives fungal disease, if improper use can cause more residual in fruit and fruit product.Therefore, carry out thiabendazole Monitoring Pesticide Residues in fruit, to grasp fruit pesticide residue situation, improve fruit quality and Protection of consumer healthy, there is vital role.
At present, the method such as residue detection many employings tlc, vapor-phase chromatography, high performance liquid chromatography of agricultural chemicals.The detectability of these methods is low, highly sensitive, existing relevant technical bid will definitely be for reference, but owing to needing expensive instrument, special operator, and sample pre-treatments is complicated, cost is high, the time is long, therefore can not meet fast and convenient Site Detection requirement better.
Immunologic detection method has fast, cheap, easy, special advantage, can be portable and carry out field monitoring.Overcome the shortcoming of traditional detection method.Set up immunologic detection method, particularly for the immunologic detection method of pesticide molecule, filter out efficient monoclonal antibody the most key.The present invention is the immunologic detection method realizing benzimidazoles sterilant thiabendazole in environment and agricultural-food, chemosynthesis is for the multiple haptens of thiabendazole, be combined into immunogen with carrier proteins or wrap by source, and filtering out the hybridoma cell strain FQ-B12 of the efficient monoclonal antibody can secreted for thiabendazole.And utilize the monoclonal antibody secreted by this hybridoma cell strain to establish enzyme-linked immune detection method for thiabendazole in environment and agricultural-food.
Summary of the invention
The object of this invention is to provide hybridoma cell strain FQ-B12, the monoclonal antibody of the anti-benzimidazoles sterilant thiabendazole of its secretion and application.
Object of the present invention realizes by following technical scheme:
1. haptens preparation
The synthesis of 1.1 2-(2 '-succinyl amido-4 '-thiazole) benzene imidazoles (H1)
1. thiazolamine-4-ethyl formate: take 1.216g thiocarbamide and be dissolved in 40mL acetonitrile, after basic dissolving, drips 3.12g ethyl bromide acetone (195g/mol, 2mL) wherein, and stirring at normal temperature reaction is spent the night.Insolubles is leached, filtrate is spin-dried for, toward being spin-dried in thing the water adding about 3mL, putting into refrigerator, spending the night.Adularescent spicule is separated out, and is leached, dry, obtains 1.85g product.
2. thiazolamine-4-formic acid: gained ester 1.85g (172g/mol) in 1 step is dissolved in methyl alcohol, adds NaoH wherein and be about 0.8g, stirring at normal temperature reaction is spent the night.If room temperature is lower, can heat a little.After spending the night, in reaction solution, drip 2mol/L HCl, to pH to 3-4.Now there is yellow solid to separate out, leached, dry, obtain product 0.91g.
3. 2-(2 '-amino-4 '-thiazole) benzene imidazoles: gained acid 0.91g (144g/mol) in 2 steps is added in 50mL Erlenmeyer flask, add equimolar O-Phenylene Diamine 0.6825g (108g/mol) again, add polyphosphoric acid again and be about 3mL, mixed.Put into microwave reaction in furnace.First low temperature 160w (1min+1min) makes dissolution of solid in polyphosphoric acid, then strengthens wattage to 200w, and each 2min interval is irradiated, and irradiates 4 times.After question response terminates, in reaction solution, add frozen water, regulate pH to 9-10 with 2mol/L NaoH, now solid is separated out, and is leached, dry, obtains product 0.68g.
1.2 5-succinoamino-2-(4-thiazole) benzene imidazoles (H2):
1. 4-thiazolecarboxylic acid ethyl ester: take 2g (44mmol) methane amide and add in 10mL benzene, add 1.4g (6.3mmol) P under stirring at room temperature
2o
5, at 25 DEG C, stir 3h.Be warming up to 40 DEG C, slowly drip ethyl bromide acetone 2mL (12.75mmol), be then warming up to backflow, reaction 2h.Add suitable quantity of water after reaction terminates, being neutralized to pH value by the NaOH solution of 1mol/L is 8, with benzene extraction, is spin-dried for benzene, obtains oily matter 1.7g (10.8mmol).Productive rate 84.7%.
2. 4-thiazolecarboxylic acid: gained ester 1.7g (10.8mmol) in 1 step is dissolved in methyl alcohol, adds NaOH wherein and be about 0.4g, stirring at normal temperature reaction is spent the night.If room temperature is lower, can heat a little.After spending the night, in reaction solution, drip 2mol/L HCl, to pH to 3-4.Now there is yellow solid to separate out, leached, dry, obtain product 0.98g (7.6mmol).Productive rate 70.3%.
3. 5-amino-2-(4-thiazole) benzene imidazoles: gained acid 0.98g (7.6mmol) in 2 steps is added in 50mL Erlenmeyer flask, add equimolar O-Phenylene Diamine 0.8208g (7.6mmol) again, add polyphosphoric acid again and be about 3mL, mixed.Put into microwave reaction in furnace.First low temperature 160w (1min+1min) makes dissolution of solid in polyphosphoric acid, then strengthens wattage to 200w, and each 2min interval is irradiated, and irradiates 4 times.After question response terminates, in reaction solution, add frozen water, regulate pH to 9-10 with 2mol/L NaOH, now solid is separated out, and is leached, dry, obtains product 0.492g (2.28mmol).Productive rate 30%.
4. 5-succinoamino-2-(4-thiazole) benzene imidazoles: 3 steps reaction products therefrom 0.492g (2.28mmol) are dissolved in 4mL pyridine, add Succinic anhydried 0.342g (3.42mmol) wherein, reaction backflow 24h.In reaction solution, add 2mol/L HCl, regulate pH to 3-4.Leached, dry, obtain crude product.With methyl alcohol: acetic acid second is cruel=and the eluent of 1: 5 crosses pillar, but sample is still not too pure.Finally use ethyl alcohol recrystallization, obtain sterling 0.41g (1.297mmol).Productive rate 56.8%.
1.3 4-benzene imidazolyl-2 (4-thiazole) butanic acids (H3):
1. 4-benzene imidazolyl-2 (4-thiazole) butyric acid second is cruel: 1.00g thiabendazole (5mmol), in the there-necked flask of 100mL, dissolves with 15mL DMF, at 0 DEG C, adds NaH (0.3g, 10mmol) while stirring.Add 0.4g benzyltriethylammoinium chloride, react after one hour, add 4-bromobutyrate 2.88g (12.9mmol).Under room temperature, (12h) is spent the night in reaction.Add water 80mL, adularescent precipitation generates.Filter.Dry.Survey fusing point: 45-47 DEG C.
2. 4-benzene imidazolyl-2 (4-thiazole) butyric acid: there-necked flask gained solid being placed in 100mL, add 20mL hydrochloric acid (2mol/L), be heated to 60-70 DEG C, solid dissolves gradually, reacts 3 hours.After cooling, adularescent solid is separated out.Filter, dry, fusing point: 219-222 DEG C.Weigh 1.3g.
2. the synthesis of artificial antigen
Take 0.25mmolH1 respectively, H2 and H3 and 0.25mmolNHS, be dissolved in reaction unit with 1.5mL DMSO; Take 0.25mmolEDC again, be dissolved in 1mLDMSO, EDC/DMSO is dissolved; DCC/DMF solution is slowly added drop-wise in above-mentioned reaction unit.Room-temperature seal reacts 7 hours under magnetic stirring.React whole liquid to put 4 DEG C of refrigerators and cool more than 2 hours, through 8000rpm centrifugal 5 minutes, get the active ester liquid of supernatant and join very slowly in the BSA albumen of 2mL (15mg/mL), react stirring at room temperature 4-6 hour under magnetic stirring.After question response completes, be placed in by reaction solution in dialysis tubing, in the PBS damping fluid of 0.2mol/L pH6.8,4 DEG C are stirred dialysis, within every four hours, change a dialyzate, dialyse 60 hours altogether.After dialysis terminates, the white emulsion (H1-BSA, H2-BSA and H3-BSA) in dialysis tubing is taken out packing to be stored in-20 DEG C of refrigerators.
Utilize aforesaid method to synthesize H1-OVA respectively, H2-OVA, H3-OVA are as envelope antigen.
3. mouse immune
By the female BAIB/C mouse in immunogen immune 6-8 age in week prepared, the immunogen consumption of every mouse is 100 μ g (calculating with protein content), altogether immunity five times at every turn.From the 4th time, latter one week of each immunity, caudal vein is taken a blood sample, and detects tiring and specificity of antibody by indirect non-competing ELISA method.
4. cytogamy
After mice serum tires no longer rising, get the BAIB/C mouse immunogen direct immunization of tiring the highest, spleen is got after three days, the SP2/0 myeloma cell of splenocyte and logarithmic phase prepares hybridoma by hybridoma technology, 37 DEG C, 5% CO2gas incubator is cultivated after 10-14 days, by detection fusion hole supernatant screening hybridoma;
5. filtering hybridoma
First merge hole supernatant adopts indirect non-competing ELISA method to carry out preliminary screening, after obtaining the fusion hole of tiring higher, confirms whether merge hole supernatant (antibody of hybridoma secretion) identifies thiabendazole molecule with indirect competitive ELISA method.Subclone (gradient dilution method) is carried out to the fusion hole that there is identity, repeats above-mentioned screening procedure.Through the screening in 3-5 cycle, determine to identify the hybridoma cell strain of thiabendazole molecule and the monoclonal antibody of secretion thereof.
6. the preservation of hybridoma cell strain
Involved in the present invention can secreting identifies that the hybridoma cell strain of thiabendazole monoclonal antibody is preserved in China typical culture collection center on May 30th, 2014; Preservation address, Wuhan, China Wuhan University; Deposit number CCTCC NO:C201491; Specific name: hybridoma cell strain FQ-B12.
Beneficial effect
(1) good stability of antigen and antibody: the thiabendazole antigen of this method synthesis has good stability, and-20 DEG C of refrigerators at least can be deposited and not affect its immunogenicity in 5 years.Cryopreservation of Hybridoma Cells at least keeps 3 years in liquid nitrogen, and the stability of antibody also can be relatively good, and antibody-20 DEG C of refrigerators of purifying at least can deposit 2 years.
(2) antibodies specific is high: the anti-thiabendazole monoclonal antibody secreted by hybridoma cell strain FQ-B12 provided by the invention to the cross reaction of the structural similitude agricultural chemicals such as derosal, thiophanate_methyl, F-1991 all lower than 0.1%.
Embodiment
The detection of embodiment 1 thiabendazole standard substance
Adopt indirect competitive ELISA method, desk study is carried out to detection curve.Method is as follows:
Bag quilt: being buffered liquid (0.05mol/L, pH9.6) by envelope antigen (H3-OVA) dilution with bag is 5 μ gmL
-1add enzyme plate, every hole 50 μ l, hatches 2h in 37 DEG C of incubators;
(2) plate is washed: wash 5 times with washings PBST (0.05% polysorbas20,0.01mol/L, pH7.4), thieving paper pats dry;
(3) close: every hole adds 200 μ L1% gelatin, hatches 1.5h for 37 DEG C;
(4) plate is washed: with (2);
(5) agricultural chemicals and mixtures of antibodies is added: with PBST phosphoric acid buffer 3 times of stepwise dilution thiabendazole standardized solution, every hole adds 25 μ L, then adds the antibody-solutions 25 μ L of dilution certain multiple, hatches 1h for 37 DEG C, PBST washs 5 times, and be arranged in parallel positive control and negative control;
(6) plate is washed: with (2);
(7) ELIAS secondary antibody body is added: every hole adds horseradish peroxidase-goat anti-mouse igg (Wuhan doctor's moral company) that 50 μ L dilute through 1: 20000 times of PBST, hatches 1h for 37 DEG C;
(8) plate is washed: with (2);
(9) develop the color: every hole adds the freshly prepared nitrite ion of 50 μ L, 37 DEG C of incubation 10min;
(10) stop: every hole adds the H of 25 μ L2mol/L
2sO
4solution;
Measure each hole light absorption value at 450nm wavelength place by microplate reader (Thermo CO.), and calculate inhibiting rate.The IC of antibody
50=6.0ng/mL.Result shows, the antibody capable of preparation identifies thiabendazole preferably.
Thiabendazole residual quantity is detected in embodiment 2 apple, pears sample
H3-OVA coated elisa plate 50 μ L/ hole, thiabendazole monoclonal antibody dilutes 1600 times as working concentration, and 1% gelatin is closure, and PBST phosphoric acid buffer pH value is 7.5, ionic strength is 0.8M, organic solvent is methyl alcohol, 50 μ L/ holes in enzyme plate.
Testing sample prepares:
Apple, pears sample are bought from Formocarbam supermarket, Nanjing, take the sample of 10g homogenate in advance, add the final concentration of standard substance (thiabendazole) to 200ng/g, mixing, room temperature leaves standstill 1h, adds the methyl alcohol of 10mL, fully vibrates, 4000rpm, centrifugal 5min, supernatant liquor moves in 25mL volumetric flask.Precipitation rejoins 10mL methyl alcohol, fully vibrates, 4000rpm, centrifugal 5min, and supernatant liquor moves in 25mL volumetric flask, and is settled to 25mL with optimum working buffer liquid (not containing organic solvent).Get the optimum working buffer liquid of sample liquid (not containing the organic solvent) dilution after part constant volume, detect for ic-ELISA.
Adopt above-mentioned ELISA detection method to detect above sample, operate as follows:
Bag quilt: be buffered liquid (0.05mol/L, pH9.6) with bag and envelope antigen (H3-OVA) is diluted 5 μ gmL
-1add enzyme plate, every hole 50 μ l, hatches 2h in 37 DEG C of incubators;
(2) plate is washed: wash 5 times with washings PBST (0.05% polysorbas20,0.01mol/L, pH7.4), thieving paper pats dry;
(3) close: every hole adds 120 μ L1% gelatin, hatches 1.5h for 37 DEG C;
(4) plate is washed: with (2);
(5) agricultural chemicals and mixtures of antibodies is added: the apple sample solution that with the addition of thiabendazole with the PBST phosphoric acid buffer 8 times of dilutions optimized, every hole adds 25 μ L, add the antibody-solutions 25 μ L of dilution certain multiple again, hatch 1h for 37 DEG C, PBST washs 5 times, and be arranged in parallel positive control and negative control;
(6) plate is washed: with (2);
(7) ELIAS secondary antibody body is added: every hole adds horseradish peroxidase-goat anti-mouse igg (Wuhan doctor's moral company) that 50 μ L dilute through 1: 20000 times of PBST, hatches 1h for 37 DEG C;
(8) plate is washed: with (2);
(9) develop the color: every hole adds the freshly prepared nitrite ion of 50 μ L, 37 DEG C of incubation 10min;
(10) stop: every hole adds the H of 25 μ L2mol/L
2sO
4solution;
In microplate reader, 450nm place reads light absorption value, positive value B0=0.998, testing sample light absorption value B=0.477.
11) data processing
Trying to achieve thiabendazole residual quantity in testing sample is C=192.2ng/g, and theoretical add value is 200ng/g, and obtaining the rate of recovery is 96.1%.
Claims (2)
1., for detecting a monoclonal antibody for benzimidazoles sterilant thiabendazole, the hybridoma cell strain FQ-B12 being CCTCC NO:C201491 by deposit number produces.
2. produce the hybridoma cell strain FQ-B12 of the monoclonal antibody for detecting benzimidazoles sterilant thiabendazole according to claim 1, be preserved in China typical culture collection center on May 30th, 2014, deposit number is CCTCC NO:C201491.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410497912.4A CN104292335A (en) | 2014-09-24 | 2014-09-24 | Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410497912.4A CN104292335A (en) | 2014-09-24 | 2014-09-24 | Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104292335A true CN104292335A (en) | 2015-01-21 |
Family
ID=52312310
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410497912.4A Pending CN104292335A (en) | 2014-09-24 | 2014-09-24 | Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104292335A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106124766A (en) * | 2016-07-05 | 2016-11-16 | 天津师范大学 | Use the method for carbendazim content in carbendazim detection of specific antibody edible fungi |
| CN109022366A (en) * | 2018-08-16 | 2018-12-18 | 江南大学 | One plant of hybridoma cell strain for secreting anti-levamisol monoclonal antibody and its application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102675463A (en) * | 2011-03-10 | 2012-09-19 | 中华人民共和国北京出入境检验检疫局 | Carbendazim monoclonal antibody, preparation method and application thereof |
| CN104017070A (en) * | 2014-06-26 | 2014-09-03 | 江南大学 | Synthesis method of high-sensitivity carbendazol complete antigen |
-
2014
- 2014-09-24 CN CN201410497912.4A patent/CN104292335A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102675463A (en) * | 2011-03-10 | 2012-09-19 | 中华人民共和国北京出入境检验检疫局 | Carbendazim monoclonal antibody, preparation method and application thereof |
| CN104017070A (en) * | 2014-06-26 | 2014-09-03 | 江南大学 | Synthesis method of high-sensitivity carbendazol complete antigen |
Non-Patent Citations (3)
| Title |
|---|
| BRANDON DL.等: "Monoclonal Antibody-Based ELISA for Thiabendazole in Liver>", 《J. AGRIC. FOOD CHEM》 * |
| HORNE E.等: "Immobilisation of thiabendazole specific antibodies on an agarose matrix for application in immunoaffinity chromatography", 《ANALYST》 * |
| 陈红平等: "农药残留免疫分析技术应用与质量控制", 《农药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106124766A (en) * | 2016-07-05 | 2016-11-16 | 天津师范大学 | Use the method for carbendazim content in carbendazim detection of specific antibody edible fungi |
| CN109022366A (en) * | 2018-08-16 | 2018-12-18 | 江南大学 | One plant of hybridoma cell strain for secreting anti-levamisol monoclonal antibody and its application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105424939B (en) | A kind of test strips for detecting carbendazim and its preparation method and application | |
| CN102161662B (en) | Artificially synthesized antigen of sildenafil and derivatives thereof as well as preparation method and application of antigen | |
| CN101865924B (en) | Method for detecting fumonisins based on immuno-magnetic bead combined enzyme-linked immunosorbent assay | |
| CN104502554B (en) | A kind of nano colloid gold label immunoassay method of tadalafil and the like | |
| CN102426229B (en) | Preparation method of human IgG immunomagnetic bead for staphylococcus aureus enrichment and application | |
| CN103941000B (en) | A kind of detect sulfa drugs and the test strips of FQNS and method | |
| CN105675874A (en) | Colloidal gold test strip for detection of imidacloprid and application thereof | |
| CN104849461A (en) | Test paper strip for detecting zearalenone and application of test paper strip | |
| CN105137009A (en) | ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit | |
| CN104327183A (en) | Preparation method and application of paclobutrazol artificial antigen and polyclonal antibody | |
| CN101830980B (en) | Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method | |
| CN105759043A (en) | Test paper for simultaneous detection of carbendazim and thiophanate-methyl residues in crops, and application and preparation method thereof | |
| CN104292335A (en) | Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12 | |
| CN109239336A (en) | A kind of test strips and its application detecting Mobucin | |
| CN109265404A (en) | A kind of preparation method and application of carbendazim haptens and antigen | |
| CN105785011A (en) | Test strip for detecting maleic hydrazide, as well as preparation method and application thereof | |
| CN103601807B (en) | The preparation method of anti-oxime carbamate class agricultural chemicals butocarboxim monoclonal antibody | |
| CN103777015B (en) | A kind of colloidal gold strip detecting erythromycin and method | |
| CN103833845B (en) | The synthetic method of a kind of m-tetrachlorophthalodinitrile artificial antigen | |
| CN103513035A (en) | Test strip and method for detecting aflatoxin M1 | |
| CN101241133A (en) | ELISA kit for detecting salbutamolum residue and method of use thereof | |
| CN105004844A (en) | Gentamicin residue diagnosis strip and application thereof | |
| CN101747429A (en) | Specific antibody against pesticide meta-tolyl-N-methylcarbamate | |
| CN109061154A (en) | A kind of fluorescent micro-ball immune chromatography test paper strip and its preparation method and application detecting metalaxyl | |
| CN107805241B (en) | Imidacloprid hapten, complete antigen, preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150121 |
|
| WD01 | Invention patent application deemed withdrawn after publication |