CN103102415A - Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl - Google Patents
Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl Download PDFInfo
- Publication number
- CN103102415A CN103102415A CN2012102383216A CN201210238321A CN103102415A CN 103102415 A CN103102415 A CN 103102415A CN 2012102383216 A CN2012102383216 A CN 2012102383216A CN 201210238321 A CN201210238321 A CN 201210238321A CN 103102415 A CN103102415 A CN 103102415A
- Authority
- CN
- China
- Prior art keywords
- parathion
- methyl
- antibody
- hybridoma
- cell
- 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
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a bispecific monoclonai antibody capable of simultaneously recognizing imidacloprid and parathion-methyl pesticides, and a preparation method thereof, belonging to the field of biological technology. The bispecific monoclonai antibody FQ-D6 which can be stably secreted and simultaneously resist imidacloprid and parathion-methyl pesticides is prepared by fusing hybridoma cell capable of secreting anti-parathion-methyl monoclonal antibody and an imidacloprid antigen-immunized rat splenocyte cell through a hydridizatio-hybridoma technique. Proved by effectiveness verification, the antibody can be used for fast and sensitively detecting residuals of the parathion-methyl and the imidacloprid in agricultural production environment and agricultural products. The preparation technology of the bispecific monoclonai antibody capable of simultaneously resisting imidacloprid and parathion-methyl pesticides is simple, convenient and feasible, the whole preparation process of the antibody is not required for special instruments and equipment, thereby being easy for factorization scale production.
Description
(1) technical field
The present invention relates to be applied to detect simultaneously Provado and parathion-methyl bispecific monoclonal antibody and preparation method, belong to biological technical field.Be exclusively used in Provado in agricultural-food and agriculture production environment, residual sensitivity, the rapid detection of parathion-methyl, be specially adapted to batch samples and detect.
(2) background technology
Provado (imidacloprid, 1-(6-chloro-3-pyridylmethyl)-N-nitro-2-tetrahydroglyoxaline imines) and parathion-methyl (Parathion-methyl, O, O-dimethyl-O-(4-nitrophenyl) thiophosphatephosphorothioate) be the sterilant that is widely used in agriculture production, Provado belongs to the moderate toxicity novel nicotinamide insecticide, and parathion-methyl belongs to high malicious organophosphate insecticides.What at present, the residue detection in agricultural-food adopted to these two kinds of agricultural chemicals is vapor-phase chromatography (GC) and high performance liquid chromatography (HPLC); These two kinds of pesticide residue immune analysis methods also there is relevant report, but simultaneously has not yet to see report for immune many method for detecting residue of these two kinds of agricultural chemicals.
The pesticide multi-residues immuno analytical method mainly contains following three kinds: 1. with on multiple hapten conjugation to a carrier proteins, immune animal obtains antiserum(antisera) and carries out the pesticide multi-residues analysis; 2. extract the public structure of a series of agricultural chemicals, design universal hapten, immune animal obtains monoclonal antibody and is used for the pesticide multi-residues analysis; 3. bi-specific antibody method.First method makes it be subject to certain limitation on producing because being merely able to obtain polyclonal antibody; The prepared monoclonal antibody of second method is merely able to identify a class agricultural chemicals, can not detect simultaneously for different types of agricultural chemicals; And bi-specific antibody has solved this problem well, and bi-specific antibody has two different antigen binding sites because of it, and reaches the purpose that detects simultaneously the different sorts agricultural chemicals.
The preparation bi-specific antibody mainly contains following three kinds of methods: chemical crosslink technique, hybrid-hybridoma technique and genetic engineering antibody technology.The present invention utilizes parathion-methyl hybridoma and the technology that the BALB/C mice splenocyte that is subjected to the Provado immunogen immune merges, and prepares the secretion trisome hybridoma of anti-Provado-parathion-methyl monoclonal antibody simultaneously.
(3) summary of the invention
Technical problem the purpose of this invention is to provide a kind of can be used for detecting simultaneously parathion-methyl and the residual bispecific monoclonal antibody of imidacloprid pesticide and preparation method thereof.Prepare by the hybridoma that will secrete anti-parathion-methyl monoclonal antibody and the strategy that the BALB/C mice splenocyte that is subjected to the Provado immunogen immune merges the trisome hybridoma that to secrete anti-parathion-methyl-Provado bispecific antibody.
Technical scheme
1. anti-parathion-methyl and Provado bispecific monoclonal antibody, produced by trisome hybridoma cell strain FQ-D6, is the IgG2b-IgG1 type, and light chain of antibody is the kappa chain.
2. the preparation method of anti-parathion-methyl and Provado bispecific monoclonal antibody comprises:
(1) the HGPRT deficient hybridoma cell strain screening of the anti-parathion-methyl monoclonal antibody of secretion
The hybridoma cell strain of the anti-parathion-methyl antibody of secretion is placed in the 5ug/ml8-AG substratum and cultivates, improve gradually 8-AG concentration in substratum in culturing process, cell strain can be adapted in the substratum of the 8-AG of 50ug/ml grow; Utilize afterwards HAT selective d MEM substratum to screen, whether survive the enzyme in identification of cell whether to lack by cell, clone under deletion condition and enlarged culturing, and cell cryopreservation is standby.
(2) be subjected to the preparation of Provado immunogen immune BALB/C mice splenocyte
Exempted from by Provado immunogen 5, serum titer reached 1: 32000 above BALB/C mice booster immunization after 3 days, and eye socket is got blood and prepared serum, the positive control during as the trisome filtering hybridoma; Draw the neck dislocation to put to death the mouse after bloodletting, and be in 75% alcohol body surface sterilization 10 minutes, move into subsequently in Bechtop, be positioned in sterile petri dish, left abdomen up; Utilize aseptic operation tweezer and eye scissors to take out mouse spleen, wash gently with the DMEM basic medium, peel off as far as possible spleen fat and reticular tissue on every side; Spleen is placed in 120 purpose nylon filter bag, and puts into the culture dish that another fills the DMEM basic culture solution; Spleen in nylon filter bag is punctured and constantly extruding, splenocyte is discharged in the DMEM basic culture solution fully; Splenocyte suspension is transferred in the 50ml centrifuge tube, and centrifugal 10 molecules of 1000rpm are abandoned supernatant, will precipitate to wash once with about 30mlDMEM nutrient solution to suspend afterwards again, and are standby
(3) cytogamy
The HGPRT deficient hybridoma cell strain of splenocyte and the good anti-parathion-methyl antibody of secretion of growth conditions is prepared the trisome hybridoma by hybrid-hybridoma technique.And in 37 ℃, volume ratio 5% CO2gas incubator is cultivated, and detects by non-competing ELISA afterwards until 7 days and merges hole supernatant screening hybridoma.
(4) screening of trisome hybridoma
Adopt indirect non-competing ELISA method to carry out preliminary screening to merging the porocyte supernatant, method is:
Dilute respectively parathion-methyl and Provado coating antigen with the coated damping fluid of carbonate, in 96 hole enzyme plates, every hole adds 50ul, in 4 ℃ of lower overnight incubation, with coating buffer in enzyme plate to the greatest extent, and washs 3 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20; Gelatin is configured to 1% confining liquid with ultrapure water, in 96 hole enzyme plates, every hole adds 200ul, hatches 1.5 hours for 37 ℃, with confining liquid in enzyme plate to the greatest extent, and washs 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20
In 96 hole enzyme plates, every hole adds the fusion porocyte supernatant to be measured that 50ul collects; To dilute with the phosphate buffered saline buffer that contains 0.05% polysorbas20 by immune serum, in 96 hole enzyme plates, every hole adds 50ul as positive control; In 96 hole enzyme plates every hole add 50ul not the fused cell supernatant as negative control.Hatched 1 hour for 37 ℃, with reaction solution in enzyme plate to the greatest extent, and wash 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Goat anti-mouse antibody with the phosphate buffered saline buffer dilution horseradish peroxidase-labeled that contains 0.05% polysorbas20, in 96 hole enzyme plates, every hole adds 50ul, hatched under 37 ℃ 45 minutes, and with reaction solution in enzyme plate to the greatest extent, and washed 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Now join the tetramethyl benzidine nitrite ion with the Trisodium Citrate substrate buffer solution, in 96 hole enzyme plates, every hole adds 50ul, and under 37 ℃ of dark conditions, reaction is 10 minutes;
In 96 hole enzyme plates, every hole adds 2mol/L sulfuric acid 25ul, color development stopping;
Enzyme plate after color development stopping is measured the single wavelength light absorption value in each hole immediately under 450nm;
Under two kinds of coating antigens, have the supernatant liquor reading of fused cell to be equal to or during greater than 2.1 times of negative control reading, such fusion hole is positive hole;
In order to determine that further screening the secreted antibody in positive hole is the bispecific monoclonal antibody of identifying simultaneously parathion-methyl and Provado, then by the screening of indirect competitive ELISA method, method is:
Dilute respectively parathion-methyl, Provado coating antigen with the coated damping fluid of carbonate, in 96 hole enzyme plates, every hole adds 50ul, in 4 ℃ of lower overnight incubation, with coating buffer in enzyme plate to the greatest extent, and washs 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Gelatin is configured to 1% confining liquid with ultrapure water, in 96 hole enzyme plates, every hole adds 200ul, hatches 1.5 hours for 37 ℃, with confining liquid in enzyme plate to the greatest extent, and washs 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Configure respectively parathion-methyl and the imidacloprid pesticide standard substance methanol solution of a series of concentration, after 20 times of the phosphate buffered saline buffer dilutions that contains 0.05% polysorbas20, fully mix with the cell culture supernatant equal-volume; Two kinds of agricultural chemicals-cell conditioned medium mixed solution is added in enzyme plate with every hole 50ul; Think the fused cell supernatant and contain 0.05% polysorbas20 phosphate buffered saline buffer equal-volume mixed solution as negative control; Hatched 1 hour for 37 ℃, with reaction solution in enzyme plate to the greatest extent, and wash 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Goat anti-mouse antibody with the phosphate buffered saline buffer dilution horseradish peroxidase-labeled that contains 0.05% polysorbas20, in 96 hole enzyme plates, every hole adds 50ul, hatched under 37 ℃ 45 minutes, and with reaction solution in enzyme plate to the greatest extent, and washed 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
Now join the tetramethyl benzidine nitrite ion with the Trisodium Citrate substrate buffer solution, in 96 hole enzyme plates, every hole adds 50ul, and under 37 ℃ of dark conditions, reaction is 10 minutes;
In 96 hole enzyme plates, every hole adds 2mol/L sulfuric acid 25ul, color development stopping; After color development stopping, enzyme plate is measured the single wavelength light absorption value in each hole immediately under 450nm.Under setting parathion-methyl coating antigen was coated, the reading that does not contain the cell conditioned medium of agricultural chemicals was B
01, the cell conditioned medium reading that contains the parathion-methyl agricultural chemicals is B
11, the cell conditioned medium liquid reading that contains imidacloprid pesticide is B
12Under the Provado coating antigen was coated, the reading that does not contain the cell conditioned medium of agricultural chemicals was B
02, the cell conditioned medium reading that contains the parathion-methyl agricultural chemicals is B
21, the cell conditioned medium liquid reading that contains imidacloprid pesticide is B
22When following phenomenon occurring simultaneously: B
11＜B
01, B
12=B
01, B
21＜B
02, B
22=B
02, can assert that just the antibody of its secretion is bispecific monoclonal antibody, and utilize limiting dilution assay to carry out subclone to cell, the supernatant of its secretion is made bispecific monoclonal antibody.
The hybridoma cell strain Classification And Nomenclature that produces above-mentioned bispecific monoclonal antibody is: hybridoma cell strain FQ-D6; On July 10th, 2012 was preserved in Chinese Typical Representative culture collection center (China, Wuhan, Wuhan University), and deposit number is: CCTCC NO:C201274
Beneficial effect
Practicality is good: traditional analysis of agricultural drugs is mainly carried out in the laboratory, its pretreatment process is loaded down with trivial details, analysis speed is slow, cost is high, the a large amount of organic solvents that use simultaneously easily cause environmental pollution, and instrumentation degree and personnel specialty competency profiling to the laboratory are higher, can not satisfy in Agricultural Products Trade requirements quick, easy, accurate, a large amount of detections.And that ELISA (Enzyme Linked Immunoadsorbent Assay) method that we provide take antigen antibody reaction as the basis has is easy and simple to handle, quick, analysis cost is low, the advantage such as safe and reliable, generally do not need expensive instrument, can simplify in a large number sample pretreatment process, professional technique to the user of service is less demanding, easily universal and popularization, be specially adapted to pattern detection in enormous quantities and Site Detection.
Sensing range is wide: utilize the bispecific monoclonal antibody of hybrid-hybridoma technique preparation, have the characteristic that detects simultaneously two class pesticide structures, compare conventional monoclonal antibody, sensing range is wider, and application prospect is better.
The antibody good stability: the hybridoma that this method obtains can be preserved 3 years under the liquid nitrogen condition at least, and purified antibody can be deposited 2 years under-20 ℃ of conditions
Description of drawings
(1) standard working curve of Provado, parathion-methyl
Embodiment
Provado, parathion-methyl residues detection in embodiment 1 tap water sample
ELISA reaction system underlying condition
Provado coating antigen concentration is 4.88ug/ml, and the ascites extension rate is 2000 times; Parathion-methyl coating antigen concentration is 2.3ug/ml, and the ascites extension rate is 4000 times.1% gelatin is made confining liquid, and methanol content is 5% in antigen antibody reaction liquid (phosphate buffered saline buffer that contains 0.05% polysorbas20), the pH value is 8.5, ionic strength is 0.8mol/L, every hole 50ul in the trace analysis plate.
Testing sample is prepared
The tap water sample: Provado and parathion-methyl agricultural chemicals are made into the methanol solution of different concns, Provado concentration is respectively 6,12ug/ml, and parathion-methyl concentration is respectively 0.64,1.28ug/ml.After above agricultural chemicals methanol solution was diluted 20 times with tap water, pesticide concentration was respectively: Provado 300,600ng/ml, parathion-methyl 32,64ng/ml.
Adopt the ELISA detection method that above sample is detected, operate as follows:
(1) coated
Provado coating antigen concentration is 4.88ug/ml, and parathion-methyl coating antigen concentration is 2.3ug/ml, and every hole adds 50ul in 96 hole enzyme plates, 4 ℃ of overnight incubation, to the greatest extent,
(2) sealing
1% gelatin is made closure, and in 96 hole enzyme plates, every hole adds 200ul, hatches 1.5 hours for 37 ℃, to the greatest extent, washs 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20, and pats dry;
(3) application of sample
Be to add in enzyme plate after the secreted antibody-solutions equal-volume of CCTCC NO:C201274 trisome hybridoma mixes Provado, the parathion-methyl solution of above-mentioned preparation with deposit number, in 96 hole enzyme plates, every hole adds 50ul, hatched 1 hour for 37 ℃, reaction solution in enzyme plate is most, and wash 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
(4) add ELIAS secondary antibody
Goat anti-mouse antibody with the phosphate buffered saline buffer dilution horseradish peroxidase-labeled that contains 0.05% polysorbas20, in 96 hole enzyme plates, every hole adds 50ul, hatched under 37 ℃ 45 minutes, and with reaction solution in enzyme plate to the greatest extent, and washed 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
(5) data processing
With the combination rate (B/B that obtains
0) in substitution Fig. 1 in the typical curve of Provado, parathion-methyl, obtain the concentration of Provado and parathion-methyl, the concentration data that obtains be multiply by multiple 2.And compare with adding concentration, obtaining the interpolation rate of recovery of Provado in tap water is 97%-104%, the interpolation rate of recovery of parathion-methyl is 87%-99%.
Provado, parathion-methyl residues detection in embodiment 2 pond water samples
ELISA reaction system underlying condition is with embodiment 1
Testing sample is prepared
The pond water sample: the pond water of collecting is centrifugal, get supernatant liquor and test.Provado and parathion-methyl agricultural chemicals are made into the methanol solution of different concns, and Provado concentration is respectively 6,12ug/ml, and parathion-methyl concentration is respectively 0.64,1.28ug/ml.After above agricultural chemicals methanol solution was diluted 20 times with pond water, pesticide concentration was respectively: Provado 300,600ng/ml, parathion-methyl 32,64ng/ml.
Adopt the ELISA detection method that above sample is detected, operate as follows:
Step (1), (2) are with embodiment 1
(3) application of sample
Add in enzyme plate with Provado, the parathion-methyl solution of above-mentioned preparation and after the antibody-solutions equal-volume mixes, in 96 hole enzyme plates, every hole adds 50ul, hatched 1 hour for 37 ℃, with reaction solution in enzyme plate to the greatest extent, and wash 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
(4) add ELIAS secondary antibody
Goat anti-mouse antibody with the phosphate buffered saline buffer dilution horseradish peroxidase-labeled that contains 0.05% polysorbas20, in 96 hole enzyme plates, every hole adds 50ul, hatched under 37 ℃ 45 minutes, and with reaction solution in enzyme plate to the greatest extent, and washed 5 times with the phosphate buffered saline buffer that contains 0.05% polysorbas20;
(5) data processing
With the combination rate (B/B that obtains
0) in substitution Fig. 1 in the typical curve of Provado, parathion-methyl, obtain the concentration of Provado and parathion-methyl, and the concentration numbers that obtains is on duty with multiple 2.And compare with adding concentration, obtaining the interpolation rate of recovery of Provado in pond water is 102%-104%, the interpolation rate of recovery of parathion-methyl is 102%-104%.
Claims (4)
1. described bispecific monoclonal antibody, be that deposit number is the trisome hybridoma cell strain FQ-D6 of CCTCC NO:C201274.
2. the secreted monoclonal antibody of the described trisome hybridoma cell strain of claim 1 FQ-D6, be the bispecific monoclonal antibody that detects simultaneously parathion-methyl and Provado, is the IgG2b-IgG1 type, and light chain of antibody is the kappa chain.
3. the described trisome hybridoma cell strain of claim 1 FQ-D6 is that the hybridoma by will secrete anti-parathion-methyl monoclonal antibody and merging through the BALB/C mice splenocyte of Provado antigen immune obtains; Concrete preparation method comprises
(1) screening of the anti-parathion-methyl hybridoma cell strain of HGPRT deficient
The hybridoma cell strain of the anti-parathion-methyl antibody of secretion is placed in 5ug/ml 8-AG substratum and cultivates, improve gradually 8-AG concentration in substratum in culturing process, cell strain can be adapted in the substratum of the 8-AG of 50ug/ml grow; Utilize afterwards HAT selective d MEM substratum to screen, whether survive the enzyme in identification of cell whether to lack by cell, clone under deletion condition and enlarged culturing, and cell cryopreservation is standby.
(2) cytogamy
Utilize indirect non-competing ELISA to detect the BALB/C mice serum that crossed by the Provado immunogen immune, get the high person of tiring and dissect and obtain spleen, the HGPRT deficient hybridoma cell strain of splenocyte and the good anti-parathion-methyl antibody of secretion of growth conditions is prepared the trisome hybridoma by hybrid-hybridoma technique.And in 37 ℃, volume ratio 5% CO2gas incubator is cultivated, and detects by non-competing ELISA afterwards until 7 days and merges hole supernatant screening hybridoma.
(3) trisome filtering hybridoma
Adopt indirect non-competing ELISA method to carry out preliminary screening to merging the porocyte supernatant: under two kinds of coating antigens of parathion-methyl and Provado, have the supernatant liquor reading of fused cell to be equal to or during greater than 2.1 times of negative control reading, such fusion hole is positive hole;
In order to determine that further screening the secreted antibody in positive hole is the bispecific monoclonal antibody of identifying simultaneously parathion-methyl and Provado, again by the screening of indirect competitive ELISA method: under setting parathion-methyl coating antigen was coated, the reading that does not contain the cell conditioned medium of agricultural chemicals was B
01, the cell conditioned medium reading that contains the parathion-methyl agricultural chemicals is B
11, the cell conditioned medium liquid reading that contains imidacloprid pesticide is B
12Under the Provado coating antigen was coated, the reading that does not contain the cell conditioned medium of agricultural chemicals was B
02, the cell conditioned medium reading that contains the parathion-methyl agricultural chemicals is B
21, the cell conditioned medium liquid reading that contains imidacloprid pesticide is B
22When following phenomenon occurring simultaneously: B
11＜B
01, B
12=B
01, B
21＜B
02, B
22=B
02, can assert that just the antibody of its secretion is bispecific monoclonal antibody, and utilize limiting dilution assay to carry out subclone to cell, the supernatant of its secretion is made bispecific monoclonal antibody.
4. produce the described trisome hybridoma cell strain of claim 1 or 2 FQ-D6 and be preserved in Chinese Typical Representative culture collection center on July 10th, 2012, deposit number is CCTCC NO:C201274.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210238321.6A CN103102415B (en) | 2012-07-11 | 2012-07-11 | Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210238321.6A CN103102415B (en) | 2012-07-11 | 2012-07-11 | Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103102415A true CN103102415A (en) | 2013-05-15 |
| CN103102415B CN103102415B (en) | 2014-07-30 |
Family
ID=48310640
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210238321.6A Active CN103102415B (en) | 2012-07-11 | 2012-07-11 | Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103102415B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103308697A (en) * | 2013-06-09 | 2013-09-18 | 卫生部北京医院 | Kit used for detecting natural bispecific antibody resistant to HCV (hepatitis C virus) coded non-structural proteins NS3 and NS5 |
| CN107305213A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | A kind of method and kit for detecting organophosphate and carbamate pesticide |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2869868Y (en) * | 2005-12-06 | 2007-02-14 | 万积成 | Colloid gold test-paper for quick detecting parathion, methyl parathion residue |
| CN101880325A (en) * | 2010-06-22 | 2010-11-10 | 南京农业大学 | Monoclonal antibody for detecting imidacloprid pesticide residue |
| CN102507930A (en) * | 2011-11-07 | 2012-06-20 | 石洪波 | Kit for quickly detecting trace imidacloprid residue |
-
2012
- 2012-07-11 CN CN201210238321.6A patent/CN103102415B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2869868Y (en) * | 2005-12-06 | 2007-02-14 | 万积成 | Colloid gold test-paper for quick detecting parathion, methyl parathion residue |
| CN101880325A (en) * | 2010-06-22 | 2010-11-10 | 南京农业大学 | Monoclonal antibody for detecting imidacloprid pesticide residue |
| CN102507930A (en) * | 2011-11-07 | 2012-06-20 | 石洪波 | Kit for quickly detecting trace imidacloprid residue |
Non-Patent Citations (2)
| Title |
|---|
| 李刚等: "抗吡虫啉单克隆抗体的制备及鉴定", 《生物工程学报》, vol. 27, no. 6, 25 June 2011 (2011-06-25), pages 943 - 951 * |
| 王刚垛等: "甲基对硫磷单克隆抗体的研制及鉴定", 《中国劳动卫生职业病杂志》, vol. 19, no. 4, 31 August 2001 (2001-08-31), pages 265 - 267 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103308697A (en) * | 2013-06-09 | 2013-09-18 | 卫生部北京医院 | Kit used for detecting natural bispecific antibody resistant to HCV (hepatitis C virus) coded non-structural proteins NS3 and NS5 |
| CN107305213A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | A kind of method and kit for detecting organophosphate and carbamate pesticide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103102415B (en) | 2014-07-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101813697A (en) | Broad-spectrum pesticide residue immunity test strip and preparation method and application thereof | |
| CN101880325A (en) | Monoclonal antibody for detecting imidacloprid pesticide residue | |
| CN104263701A (en) | Chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof | |
| CN102994445A (en) | Method for performing screening in virus-sensitive cell cloning by applying indirect immunofluorescence assay technology | |
| CN105400742A (en) | Hybridoma cell strain IMI-G12, neonicotinoid pesticide universal monoclonal antibody generated by same and application | |
| CN101519447A (en) | Anti-rabbit hemorrhagic disease virus VP60 albumen monoclonal antibody | |
| CN104004717A (en) | Anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof | |
| CN104327186A (en) | Anti-bifenthrin monoclonal antibody and application thereof | |
| CN108251381A (en) | A kind of paraquat monoclonal antibody hybridoma cell strain and its application | |
| CN102994455B (en) | Monoclonal antibody and kit for cucumber green mottle mosaic viruses (CGMMVs) | |
| CN104849251A (en) | Time resolution fluorescence immunoassay method and kit for fast detecting gutter oil | |
| CN103102415B (en) | Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl | |
| CN104974256A (en) | Anti-thiacloprid monoclonal antibody and application thereof | |
| CN101846685A (en) | Enzyme-linked immunoassay kit for chlamys ferreri blood cells and preparation method thereof | |
| CN101975858A (en) | Verticiliumdahliae enzyme linked immunodetection kit and applications thereof | |
| CN105572369A (en) | Time-resolved fluorescent immunoassay kit for detecting glyphosate and detecting method of kit | |
| CN105319366A (en) | Time-resolved fluorescence immunoassay kit detecting methyl parathion and detection method thereof | |
| CN101709087B (en) | Puccinia triticina f.sp.tritic monoclonal antibody and application thereof | |
| CN103472228B (en) | Malachite green vestigial single step chemiluminescence enzyme immunoassay detection method and kit | |
| CN109627328A (en) | Method for preparing monoclonal antibody based on Raman spectrum and microlayer model technology | |
| CN101781656A (en) | Expression of odontoglossum ringspot virus (ORSV) coat protein gene and preparation method of antibody | |
| CN101759800B (en) | Preparation of dirhamnolipid artificial antigen and antibody, enzyme-linked immunity test paper, preparation and detection method thereof | |
| CN110244046A (en) | A kind of Citrus Huanglongbing pathogen colloidal gold colloidal gold detection test paper strip and preparation method thereof, kit | |
| CN104316688B (en) | A kind of DASELISA immunization detecting xanthomonas oryzae pv. oryzicola | |
| CN101598732A (en) | The ELISA detection method of a kind of red-tide algae toxin okadaic acid OA |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |