CN103675261A - Method for marking clenbuterol monoclonal antibody by fluorescent microsphere - Google Patents
Method for marking clenbuterol monoclonal antibody by fluorescent microsphere Download PDFInfo
- Publication number
- CN103675261A CN103675261A CN201310348210.5A CN201310348210A CN103675261A CN 103675261 A CN103675261 A CN 103675261A CN 201310348210 A CN201310348210 A CN 201310348210A CN 103675261 A CN103675261 A CN 103675261A
- Authority
- CN
- China
- Prior art keywords
- fluorescent microsphere
- biotin
- monoclonal antibody
- clenbuterol
- streptavidin
- 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
Images
Classifications
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
-
- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention belongs to the technical field of biology and discloses a method for marking a clenbuterol monoclonal antibody by a fluorescent microsphere. The method specifically comprises the following steps: coupling the amino modified fluorescent microsphere with an active esterifiable biotin to obtain a coupling matter; mixing streptavidin with the coupling matter in a ratio of 1000:1 and reacting to obtain a coupling matter of the fluorescent microsphere and biotin-streptavidin; and finally, adding the biotinylated clenbuterol monoclonal antibody into a fluorescent microsphere-biotin-streptavidin coupling matter solution and reacting to obtain a target product. The obtained novel coupling matter for marking the clenbuterol monoclonal antibody by the fluorescent microsphere can be effectively applied to a clenbuterol fluorescent microsphere test strip; correspondingly, the specificity and the sensitivity of the test strip are improved so that the residual amount of clenbuterol in animal foodstuff can be rapidly, accurately, qualitatively and quantitatively detected.
Description
Technical field
The invention belongs to β in food security
2-receptor stimulating agent residue detection field, is specifically related to a kind of conjugate with fluorescent microsphere mark clenbuterol monoclonal antibody and preparation method thereof.
Background technology
Clenbuterol (clenbuterol) is as a kind of β
2-receptor stimulating agent can play the function that regulates sympathetic activation, lax tracheal smooth muscle, and the form of Chang Yiqi hydrochloride is used, clinically can be for the treatment of humans and animals asthma class illness.But while giving detoxification for a long time with the clenobuterol hydrochloride over therapeutic dose, it has nutrition reallocation effect in animal body, can significantly improve lean meat percentage and reduce fat percentage, therefore a lot of lawless persons are often used for animal feeding using clenobuterol hydrochloride as feed addictive, thereby make meat lean meat percentage increase, go on the market ahead of time, reduce costs.The abuse of clenobuterol hydrochloride will certainly be residual in livestock products, and bring sizable threat to the mankind's health.
Immunochromatography technique owing to having fast, accurately, advantage easily, in the detection application of clenobuterol hydrochloride and substitute thereof, obtained very large popularization, it be the specific reaction of take between antigen-antibody as basis, by label, certain material is carried out the research means of qualitative or quantitative detection.At present, the label that is usually used in immunochromatography technique detection clenobuterol hydrochloride has collaurum, fluorescent microsphere etc., these labels have kept the activity of clenbuterol monoclonal antibody, and the activity of label is unaffected, after it reacts with corresponding antigens, can directly measure the label in compound, directly target substance be carried out to qualitative even quantitative test.
At present, between label and antibody, be mostly the direct coupling of mode, the particularly mark of beta-receptor agonist antibody by covalent bond or physisorption.This kind of coupling mode has coupling time advantage short, simple to operate, but this coupling mode, because antibody is combined with label and is made some antigen binding site conductively-closeds arbitrarily, so-called sterically hindered, cause the dispersiveness of antibody labeling efficiency and label to reduce.
Summary of the invention
The object of the invention is the defect for above-mentioned prior art, the method of fluorescent microsphere mark clenbuterol monoclonal antibody is provided, the method has reduced sterically hindered that antibody is connected with fluorescent microsphere, improved labeling effciency high, the conjugate making can show good colloidal stability and bonding properties.
The technical scheme that the present invention takes is to achieve these goals:
The method of fluorescent microsphere mark clenbuterol monoclonal antibody: (1) amido modified fluorescent microsphere obtains fluorescent microsphere-biotin conjugates with biotin coupling reaction 2 h in PBS solution of active esterification, more unconjugated biotin is removed in centrifugal, washing; (2) Streptavidin is reacted after 2 h with fluorescent microsphere-biotin conjugates mixing jog, Streptavidin centrifugal, remove not coupling with PBS solution washing, obtains fluorescent microsphere-biotin-Streptavidin conjugate; (3) biotinylated clenbuterol monoclonal antibody being joined to jog in the conjugate solution of fluorescent microsphere-biotin-Streptavidin reacts after 1 h, washing, centrifugal to remove the biotinylation clenbuterol monoclonal antibody in not coupling, obtaining fluorescent microsphere mark clenbuterol monoclonal antibody is fluorescent microsphere-biotin-Streptavidin-biotinylation clenbuterol monoclonal antibody; The biotin of described active esterification is for to be dissolved in biotin-PEG5000 compound in DMF solution, add dicyclohexyl carbon imines and N-hydroxy-succinamide after stirring at room react 24 h, the centrifugal precipitation of abandoning then, obtains the biotin of active esterification; The preparation method of described amido modified fluorescent microsphere: 10 mg fluorescent microspheres are cleaned rear ultrasonic dissolution in 1 mL distilled water, add after 20 μ L glacial acetic acids and 20 μ L (3-trimethoxy silicon propyl group)-diethyl ethylenediamine ultrasonic dissolution, magnetic agitation 3～4 h, also resuspended with the MES damping fluid washing of 10 mM pH5.5; Described fluorescent microsphere particle diameter is 175 nm.
Described biotinylation clenbuterol monoclonal antibody preparation method: with 1 mL DMSO solution, dissolve biotin N-hydroxy-succinamide ester NHSB and be mixed with the solution that concentration is 1 mg/mL, getting 120 μ L joins in 1mL clenbuterol monoclonal antibody solution, at room temperature continue to stir insulation 2 ~ 4 h; Then add the NH4Cl solution of 9.6 μ L 1 mol/L at room temperature to stir 10 min, with PBS solution, at 4 ℃, fully dialyse and remove free biotin; By the molecular sieve column of 1 ml on the sample after having dialysed, with the slow wash-out of PBS solution, antibody, under washing between 1 ~ 3 ml, then adds the BSA solution of Sodium azide and 1.0 g/L in the target product of collecting, and keeps in Dark Place standby at 4 ℃.
The molar ratio of described Streptavidin and fluorescent microsphere-biotin conjugates hybrid reaction is 1000:1.
The molar ratio of the conjugate of described fluorescent microsphere mark biotinylation clenbuterol monoclonal antibody and fluorescent microsphere-biotin-Streptavidin is 1:1000.
The invention has the beneficial effects as follows:
1, labeling effciency is high: compare with traditional direct coupling antibody of fluorescent microsphere, biotin-Streptavidin reduces the space steric effect of combination between fluorescent microsphere and clenbuterol monoclonal antibody as coupling arm, and a fluorescent microsphere-PEG
5000-biotin-Streptavidin can with the coupling of biotinylated antibody 1:4, thereby make labeling effciency obtain correspondingly improving.
2, dispersiveness and solubility are good: the biotin of being combined with fluorescent microsphere is to use hydrophilic PEG
5000the compound of modifying, hydrophilic PEG
5000embedding improved fluorescent microsphere-antibody coupling matter in the solubility of redissolving in liquid, and because longer coupling arm, conjugate can well be dispersed in solution, substantially flocculation sediment can not occur, and shows good colloidal stability.
3, associativity is strong: PEG
5000after the biotin of-biotin-Streptavidin coupling arm on clenbuterol monoclonal antibody is combined, guaranteed the antibody combining site directional profile on fluorescent microsphere surface, strengthened the bonding properties of fluorescent microsphere-clenbuterol monoclonal antibody conjugate and target analyte, this point is mainly reflected in sensitivity and the stability of the fluorescent microsphere test strips of raising.
Accompanying drawing explanation
Above-mentioned and/or additional aspect of the present invention and advantage accompanying drawing below combination obviously and is easily understood becoming the description of embodiment:
The fluorescent microsphere mark clenbuterol monoclonal antibody that Fig. 1 the present invention makes.
Embodiment
The present embodiment has provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Biotin-PEG
5000compound is purchased from the scorching happy biological company limited in Shanghai;
Fluorescent microsphere is purchased from German Merck company, and particle diameter is 175 nm.
embodiment 1: the preparation method of the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody in the present invention
1, the preparation method of amido modified fluorescent microsphere
10 mg fluorescent microspheres are cleaned rear ultrasonic dissolution in 1 mL distilled water, add after 20 μ L glacial acetic acids and 20 μ L (3-trimethoxy silicon propyl group)-diethyl ethylenediamine ultrasonic dissolution, magnetic agitation 3～4 h, also resuspended with MES damping fluid (pH5.5) washing of 10 mM.
, active esterification the preparation method of biotin
Accurately take 4.5 mg biotin-PEG
5000compound is dissolved in the DMF solution of 2 mL, adds after 20.5 mg dicyclohexyl carbon imines and 10.5 mg N-hydroxy-succinamides, and room temperature magnetic agitation is reacted 24 h, then, with centrifugal 5 min of 4000 rpm, abandons precipitation, obtains the biotin of active esterification, standby.
3, the method for fluorescent microsphere mark clenbuterol monoclonal antibody, it is characterized in that: described biotinylation clenbuterol monoclonal antibody preparation method: with 1 mL DMSO solution, dissolve biotin N-hydroxy-succinamide ester (NHSB) and be mixed with the solution that concentration is 1 mg/mL, getting 120 μ L joins in the clenbuterol monoclonal antibody solution that 1mL concentration is 1.7 mg/mL, at room temperature continue to stir insulation 2 ~ 4h; Then the NH that adds 9.6 μ L
4cl (1 mol/L) solution at room temperature stirs 10 min, with PBS solution, at 4 ℃, fully dialyses and removes free biotin; By the molecular sieve column of 1 ml on the sample after having dialysed, with the slow wash-out of PBS solution, antibody, under washing between 1 ~ 3 ml, then adds the BSA solution of Sodium azide and 1.0 g/L in the target product of collecting, and keeps in Dark Place standby at 4 ℃.
4, with the immune microsphere of 5 mL system mark 35 μ g/mg (antibody/microballoon), concrete grammar: the biotin of the fluorescent microsphere that 2.0 mg are amido modified and the active esterification of 3.5 mg is coupling reaction 2 h in the PBS of 5 mL solution (pH 7.4 left and right), unconjugated biotin is removed in centrifugal, washing again, obtains fluorescent microsphere-biotin conjugates; Above-mentioned fluorescent microsphere-the biotin conjugates making is joined in 10 mg/mL solution of streptavidin, the molar ratio of described Streptavidin and fluorescent microsphere-biotin conjugates hybrid reaction is 1000:1, guarantee that final volume is 5 mL, jog reacts after 2 h, Streptavidin centrifugal, remove not coupling with the washing of PBS buffer solution, obtains fluorescent microsphere-biotin-Streptavidin conjugate; Again the biotinylated clenbuterol monoclonal antibody of respective markers amount (diluting 10 times also slowly adds) being joined to above-mentioned conjugate solution to final concentration is 35 μ g/mg, jog reacts after 1 h, washing, centrifugal to remove the biotinylated antibody in not coupling, obtain the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody, with 500 μ L redissolution liquid, redissolve.
The above-mentioned novel conjugate making is compared with traditional fluorescent microsphere-antibody coupling matter, dispersed obviously better at electric Microscopic observation, under 4 ℃ of conditions, places after 6 months, still very transparent, flocculation sediment phenomenon can not occur, and shows good colloidal stability.Its stronger binding ability is improved and is embodied by the fluorescent microsphere test strips sensitivity in embodiment below.
embodiment 2: the application of the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody in fluorescent microsphere test strips
1, the preparation of fluorescent micro-ball immune chromatography test paper strip
The preparation of fluorescent microsphere pad: redissolve conjugate that embodiment 1 makes to initial volume with the PNPB (wherein comprising 5% sucrose and 0.05% Tween-20) of 0.01 M, with BIODOT Dispensing System, according to the amount of 4 μ L/cm, be sprayed on glass fibre membrane, 25 ℃ of vacuum drying 1～2 h, are put in dry environment standby.
Anti-being coated with on NC film of the anti-mouse two of Clenbuterol-BSA conjugate and donkey: be 0.4 mg/mL and 0.4 mg/mL by the PBS damping fluid adjustment kit of 0.01 M by substrate concentration respectively, spray film amount is 0.74 μ L/cm, detection line is coated with Clenbuterol-BSA conjugate, the anti-mouse two of the coated donkey of nature controlling line is anti-, two zone position, 6 mm of being separated by, nature controlling line is apart from NC film top 10 mm, and detection line is apart from NC film bottom 9 mm, after 37 ℃ of drying and processings spend the night, under the environment of drying at room temperature, save backup.
The preparation of fluorescent micro-ball immune chromatography test paper strip: paste successively filter paper, sample pad, fluorescent microsphere pad, NC film and thieving paper on base plate with connecting, the test paper batten pasting cuts into test strips with cutting machine, and be assembled in ready test paper barrel, pack aluminium foil bag into, add after drying agent, sealing is preserved, and under the environment of drying at room temperature, at least can preserve 1 year.
, fluorescent microsphere test strips quantitatively detects the Clenbuterol in pork
Sample pre-treatments: get 2.5 g pork samples in 50 mL centrifuge tubes, add 5 mL distilled water to heat in boiling water and extract 15 min, take out whole supernatants in 10 new mL centrifuge tubes, add standing 1 min after 1 mL normal hexane jog, remove fat deposit above, subnatant is standby.
Detecting step: (1) adds the sample extraction liquid of negative sample and interpolation series concentration (0.5,1,2,3,4,5,6,8,10,15 ng/mL) in test strips sample aperture, react after 10 min, test strip is put into fluorescent microsphere and read instrument detection window;
(2) fluorescent microsphere, under LED lamp source excitation, sends strong fluorescence;
(3) fluorescence of transmitting, after CCD scanning system converges, is assembled pipe through photoelectricity, sends into photomultiplier, and light signal is enhanced, and in process signal conversion element, after software is processed, the power of fluorescence is inputted out with the height of numerical value.
(4) in experimentation, with series concentration, measure the numerical value of its corresponding fluorescence intensity, thereby set up a typical curve according to this series of values and Clenbuterol standard items corresponding concentration.Finally, according to the output numerical value that detects sample, calculate the content that detects Clenbuterol in sample.
(5) draw 100 μ L extracts, detect, finally according to the numerical value that detects the data output of sample, be respectively 4.08, look into canonical plotting and show that the residual quantity that detects Clenbuterol in sample is 0.
embodiment 3: in the fluorescent microsphere test strips of preparing with the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody, detect the Determination of Clenbuterol Residues in pork liver
Sample-pretreating method, test strips are made with embodiment 2.
Qualitative and quantitative analysis method is with embodiment 2, to add series concentration (0.5,1,2,3,4,5,6,8,10,15 ng/mL) and corresponding fluorescence intensity level drawing standard curve.Draw 100 μ L extracts and add in sample aperture, after 10 min, with reading instrument, detect, according to the numerical value that detects the data output of sample, be respectively 3.67, look into canonical plotting and draw residual quantity 0.8 ng/mL that detects Clenbuterol in sample.
embodiment 4: in the fluorescent microsphere test strips of preparing with the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody, detect the Determination of Clenbuterol Residues in feed
Sample pre-treatments: take 0.5 g Feed Sample, add 1 mL sample extracting solution (pH7.0 50 mmol/L Tris-HCl), stir 1 min in test tube, after standing 10 min, get supernatant liquor and make testing sample.
Test strips is made with embodiment 2.
Qualitative and quantitative analysis method is with embodiment 2, to add series concentration (0.1,0.5,1,2,2.5,3,5,7,10,15 ng/mL) and corresponding fluorescence intensity level drawing standard curve.Draw 100 μ L extracts and add in sample aperture, after 10 min, with reading instrument, detect, according to the numerical value that detects the data output of sample, be respectively 4.03, look into canonical plotting and show that the residual quantity that detects Clenbuterol in sample is 0.
embodiment 5: in the fluorescent microsphere test strips of preparing with the novel conjugate of fluorescent microsphere mark clenbuterol monoclonal antibody, detect the Determination of Clenbuterol Residues in pig urine
Urine sample can directly detect, and does not need pre-treatment.Qualitative and quantitative analysis method is with embodiment 2, to add series concentration (0.1,0.5,1,1.5,2,2.5,3,5,7,10 ng/mL) and corresponding fluorescence intensity level drawing standard curve.Draw 100 μ L urine samples and add in sample aperture, after 10 min, with reading instrument, detect, according to the numerical value that detects the data output of sample, be respectively 2.79, look into canonical plotting and show that the residual quantity that detects Clenbuterol in sample is 1.46 ng/mL.
Claims (4)
1. the method for fluorescent microsphere mark clenbuterol monoclonal antibody, it is characterized in that: (1) amido modified fluorescent microsphere obtains fluorescent microsphere-biotin conjugates with biotin coupling reaction 2 h in PBS solution of active esterification, more unconjugated biotin is removed in centrifugal, washing; (2) Streptavidin is reacted after 2 h with fluorescent microsphere-biotin conjugates mixing jog, Streptavidin centrifugal, remove not coupling with PBS solution washing, obtains fluorescent microsphere-biotin-Streptavidin conjugate; (3) biotinylated clenbuterol monoclonal antibody being joined to jog in the conjugate solution of fluorescent microsphere-biotin-Streptavidin reacts after 1 h, washing, centrifugal to remove the biotinylation clenbuterol monoclonal antibody in not coupling, obtaining fluorescent microsphere mark clenbuterol monoclonal antibody is fluorescent microsphere-biotin-Streptavidin-biotinylation clenbuterol monoclonal antibody;
The biotin of described active esterification is for to be dissolved in biotin-PEG5000 compound in DMF solution, add dicyclohexyl carbon imines and N-hydroxy-succinamide after stirring at room react 24 h, the centrifugal precipitation of abandoning then, obtains the biotin of active esterification; The preparation method of described amido modified fluorescent microsphere: 10 mg fluorescent microspheres are cleaned rear ultrasonic dissolution in 1 mL distilled water, add after 20 μ L glacial acetic acids and 20 μ L (3-trimethoxy silicon propyl group)-diethyl ethylenediamine ultrasonic dissolution, magnetic agitation 3～4 h, also resuspended with the MES damping fluid washing of 10 mM pH5.5; Described fluorescent microsphere particle diameter is 175 nm.
2. the method for fluorescent microsphere mark clenbuterol monoclonal antibody according to claim 1, it is characterized in that: described biotinylation clenbuterol monoclonal antibody preparation method: with 1 mL DMSO solution, dissolve biotin N-hydroxy-succinamide ester NHSB and be mixed with the solution that concentration is 1 mg/mL, getting 120 μ L, to join 1mL concentration be in 1.7 mg/mL clenbuterol monoclonal antibody solution, at room temperature continue to stir insulation 2 ~ 4 h; Then add the NH4Cl solution of 9.6 μ L 1 mol/L at room temperature to stir 10 min, with PBS solution, at 4 ℃, fully dialyse and remove free biotin; By the molecular sieve column of 1 ml on the sample after having dialysed, with the slow wash-out of PBS solution, antibody, under washing between 1 ~ 3 ml, then adds the BSA solution of Sodium azide and 1.0 g/L in the target product of collecting, and keeps in Dark Place standby at 4 ℃.
3. the method for fluorescent microsphere mark clenbuterol monoclonal antibody according to claim 1, is characterized in that: the molar ratio of described Streptavidin and fluorescent microsphere-biotin conjugates hybrid reaction is 1000:1.
4. the method for fluorescent microsphere mark clenbuterol monoclonal antibody according to claim 1, is characterized in that: the molar ratio of the conjugate of described fluorescent microsphere mark biotinylation clenbuterol monoclonal antibody and fluorescent microsphere-biotin-Streptavidin is 1:1000.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310348210.5A CN103675261A (en) | 2013-08-12 | 2013-08-12 | Method for marking clenbuterol monoclonal antibody by fluorescent microsphere |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310348210.5A CN103675261A (en) | 2013-08-12 | 2013-08-12 | Method for marking clenbuterol monoclonal antibody by fluorescent microsphere |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103675261A true CN103675261A (en) | 2014-03-26 |
Family
ID=50313447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310348210.5A Pending CN103675261A (en) | 2013-08-12 | 2013-08-12 | Method for marking clenbuterol monoclonal antibody by fluorescent microsphere |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103675261A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104530459A (en) * | 2014-12-10 | 2015-04-22 | 南京邮电大学 | Preparation method of polystyrene fluorescent microsphere coupled with antibody |
| CN106405075A (en) * | 2016-08-31 | 2017-02-15 | 上海美吉生物医药科技有限公司 | Immunomagnetic bead and preparation method thereof |
| CN109358197A (en) * | 2018-11-29 | 2019-02-19 | 中国农业科学院油料作物研究所 | A kind of fluorescence detection device and fluorescence detection method based on bridging antibody coupling fluorescent microsphere |
| CN109765384A (en) * | 2019-01-29 | 2019-05-17 | 北京勤邦生物技术有限公司 | A kind of canine coronavirus antibody fluorescence test strip and its preparation method and application |
| CN116068207A (en) * | 2023-03-24 | 2023-05-05 | 山东康华生物医疗科技股份有限公司 | Kit for detecting NK cell activity |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2650008A1 (en) * | 2006-04-25 | 2007-11-01 | Centre National De La Recherche Scientifique (Cnrs) | Functionalization of metal nanoparticles with oriented proteins |
| CN101231293A (en) * | 2008-02-14 | 2008-07-30 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Method for detecting vestigial clenbuterol |
| CN101435823A (en) * | 2007-11-12 | 2009-05-20 | 无锡中德伯尔生物技术有限公司 | Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof |
| CA2812208A1 (en) * | 2010-09-24 | 2012-03-29 | The Governing Council Of The University Of Toronto | Synthesis of fluorescent noble metal nanoparticles |
| CN103163297A (en) * | 2011-12-16 | 2013-06-19 | 王哲 | Multifunctional fluorescent immunochromatographic rapid quantitative detection card |
-
2013
- 2013-08-12 CN CN201310348210.5A patent/CN103675261A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2650008A1 (en) * | 2006-04-25 | 2007-11-01 | Centre National De La Recherche Scientifique (Cnrs) | Functionalization of metal nanoparticles with oriented proteins |
| CN101435823A (en) * | 2007-11-12 | 2009-05-20 | 无锡中德伯尔生物技术有限公司 | Fluorescent micro-ball immune chromatography test paper strip for detecting residual animal medicine and preparing method thereof |
| CN101231293A (en) * | 2008-02-14 | 2008-07-30 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Method for detecting vestigial clenbuterol |
| CA2812208A1 (en) * | 2010-09-24 | 2012-03-29 | The Governing Council Of The University Of Toronto | Synthesis of fluorescent noble metal nanoparticles |
| CN103163297A (en) * | 2011-12-16 | 2013-06-19 | 王哲 | Multifunctional fluorescent immunochromatographic rapid quantitative detection card |
Non-Patent Citations (2)
| Title |
|---|
| LIN WANG 等: "Fluorescent Nanoparticles for Multiplexed Bacteria Monitoring", 《BIOCONJUGATE CHEM.》 * |
| 朱正美 刘辉 主编: "《简明免疫学技术》", 31 July 2002, 科学出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104530459A (en) * | 2014-12-10 | 2015-04-22 | 南京邮电大学 | Preparation method of polystyrene fluorescent microsphere coupled with antibody |
| CN106405075A (en) * | 2016-08-31 | 2017-02-15 | 上海美吉生物医药科技有限公司 | Immunomagnetic bead and preparation method thereof |
| CN109358197A (en) * | 2018-11-29 | 2019-02-19 | 中国农业科学院油料作物研究所 | A kind of fluorescence detection device and fluorescence detection method based on bridging antibody coupling fluorescent microsphere |
| CN109765384A (en) * | 2019-01-29 | 2019-05-17 | 北京勤邦生物技术有限公司 | A kind of canine coronavirus antibody fluorescence test strip and its preparation method and application |
| CN109765384B (en) * | 2019-01-29 | 2022-11-18 | 北京勤邦生物技术有限公司 | Canine coronavirus antibody fluorescence detection test strip and preparation method and application thereof |
| CN116068207A (en) * | 2023-03-24 | 2023-05-05 | 山东康华生物医疗科技股份有限公司 | Kit for detecting NK cell activity |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103439490A (en) | Method of using fluorescent microsphere to mark ractopamine monoclonal antibody | |
| CN103675261A (en) | Method for marking clenbuterol monoclonal antibody by fluorescent microsphere | |
| CN103116023B (en) | ECL (electrochemiluminescence) immunosensor for detecting tumor markers and preparation method and applications thereof | |
| AU706668B2 (en) | Sulfo benz{e}indocyanine fluorescent dyes | |
| CN204188616U (en) | Procalcitonin immunochromatographiassay assay quantitative detection test paper | |
| CN104237537A (en) | Immune chromatography fluorescence reagent strip for detecting cardiac troponin and preparation method thereof | |
| CN103472221A (en) | Method for marking salbutamol polyclonal antibody by fluorescent microsphere | |
| CN104391117B (en) | Preparation method and application of gastric cancer antigen electrogenerated chemiluminescence sensor based on PP<y>-NH2GO-Ag2Se@CdSe | |
| CN110308286B (en) | Enhanced thyroglobulin electrochemiluminescence immunosensor based on photothermal release signals | |
| CN104459157B (en) | A kind of preparation method of the immunosensor based on ferrocene-cuprous oxide cubic nanometer Framework Mark and application | |
| CN105115961A (en) | Method for preparing electrochemical luminescence sensor made of nano-composites | |
| CN102169119B (en) | Preparation method for nano -gold immune probe | |
| CN104297479A (en) | Preparation method and application of electrochemiluminescence immunoassay sensor for detecting tumor marker | |
| CN104407140A (en) | Preparation method for immunosensor based on gamma-polyglutamic acid grafted dopamine and chitosan complex micelles | |
| CN102707060B (en) | Chemiluminescent test kit for testing activity of MGMT (O6-Methylguanine DNA-Methyltransferase) and test method | |
| CN104237512A (en) | Immune colloidal gold test stripe and preparation method and application | |
| CN114113582B (en) | Metal organic framework nanoenzyme biological probe and ELISA kit | |
| CN1687784A (en) | Test paper for testing afatoxin by nontoxic system immunochromatographic techniqul and preparing process thereof | |
| CN106370858A (en) | Potential addressing mode-based double tumor marker photoelectric detection method | |
| CN107561269A (en) | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol | |
| CN113970635B (en) | Immunochromatography test paper and preparation method and application thereof | |
| CN1232825C (en) | Reference immuno chromatographic test paper for detecting clenbuterol hydrochloride and detection method thereof | |
| CN102183641A (en) | Ractopamine immunochromatographic assay test paper strip | |
| CN104198706A (en) | Method for preparing electrochemical immunosensor for detecting tumor markers | |
| CN104459128A (en) | Salmonella immunochromatography test strip based on low-noise excitation type fluorescent mark |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140326 |