CN104099300A - Hybridoma cell line able to secrete anti-bovine immunoglobulin IgG monoclonal antibody and application thereof - Google Patents
Hybridoma cell line able to secrete anti-bovine immunoglobulin IgG monoclonal antibody and application thereof Download PDFInfo
- Publication number
- CN104099300A CN104099300A CN201410247248.8A CN201410247248A CN104099300A CN 104099300 A CN104099300 A CN 104099300A CN 201410247248 A CN201410247248 A CN 201410247248A CN 104099300 A CN104099300 A CN 104099300A
- Authority
- CN
- China
- Prior art keywords
- igg
- antibody
- add
- monoclonal 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
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a screening method for a hybridoma cell line able to secrete an anti-bovine immunoglobulin IgG monoclonal antibody for bovine immunoglobulin IgG detection, the hybridoma cell line, the monoclonal antibody secreted thereby and application in the field of bovine immunoglobulin IgG detection. The hybridoma cell is preserved in China Center For Type Culture Collection, Wuhan University at Luojiashan, Wuchang, Wuhan City in Hubei Province, and the preservation number is CCTCC No:2013183. The anti-bovine immunoglobulin IgG monoclonal antibody secreted by the hybridoma cell has the advantages of strong specificity, large affinity and high titer and the like, and can be widely applied in the detection reagent or detection equipment field of bovine immunoglobulin IgG. A bovine immunoglobulin IgG rapid detection card built based on the immunochromatography principle can be used for detection of bovine immunoglobulin IgG in colostrum and its products, bovine blood and other samples, and has significant advantages compared with conventional detection methods in the aspects of specificity, sensitivity and detection efficiency, etc.
Description
Technical field
The present invention relates to B-IgG IgG field of immunodetection, especially relate to a kind of hybridoma cell strain 9C6 and application thereof of secreting anti-B-IgG IgG monoclonal antibody.
Background technology
B-IgG system physico-chemical property with on Classification And Nomenclature to other Mammals and similar, mainly comprise IgG, IgA and IgM.IgG is divided into IgG1 and two subclass of IgG2, and wherein IgG1 is main component.IgG molecule is made up of 4 peptide chains, and 2 long-chains are called heavy chain (heavy chain, H), are made up of about 440 amino-acid residues, and molecular weight is 50～70kD approximately; Article 2, short chain is called light chain (light chain, L), is made up of the about 22.5kD of molecular weight about 220 amino acid.Article 4, peptide chain connects together by interchain disulfide bond (SS-).B-IgG extensively distributes in the body fluid of ox, and in colostrum, blood, content is higher.In many biologically active substances of colostrum, immunoglobulin G (IgG) is most important one.In bovine coloctrum, the content of IgG is restricted by age, the lactation period of cow and the many factors such as environment of feeding.Research shows, that immunoglobulin (Ig) has is antibacterial, regulate the multiple biological activity such as immunizing power, protection intestinal mucosa.In the exploitation of colostral milk products, the content of IgG directly determines quality and the price of product.Therefore research and develop anti-B-IgG IgG monoclonal antibody, the method for setting up rapid detection ox IgG has great importance.
The detection method of B-IgG IgG mainly comprises agar diffusion method (SRD), liquid phase chromatography (HPLC) and euzymelinked immunosorbent assay (ELISA) (ELISA) etc. at present.Wherein agar diffusion method is easy and simple to handle, and equipment is simple, the lower visual result of cost of determination.But due to diffusion diameter measuring error, cause result difference larger, accuracy and sensitivity are lower, labour intensity is large simultaneously, length consuming time.High-efficient liquid phase chromatogram technique analysis is accurate, reproducible, but needs complex instrument equipment, and sample pretreatment process is loaded down with trivial details, and testing process length consuming time is high to experimental situation requirement, is difficult to realize rapid detection.Euzymelinked immunosorbent assay (ELISA) is highly sensitive, and high specificity can be measured tens hundreds of sample even simultaneously, in research abroad application general, but this method complicated operation, length consuming time, expense is high.Immunochromatographic method has the advantages such as high specificity, highly sensitive, sample pre-treatments is simple, testing cost is low, is suitable for on-the-spot batch detection, and development in recent years is very fast.In colostrum processing and quality examination and milk cow calf are fed the production reality of management, in the urgent need to setting up the rapid free epidemiology detection method for the B-IgG IgG of colostrum, blood and urine.These fast methods all need specific recognition B-IgG IgG(to comprise IgG1 and IgG2) monoclonal antibody.Therefore, utilize the anti-B-IgG IgG(of hybridoma technology preparation secretion to comprise IgG1 and IgG2) monoclonal antibody, and research and develop on this basis B-IgG IgG rapid free epidemiology detection technique, significant.
Summary of the invention
Problem to be solved by this invention is to utilize hybridoma technology seed selection can secrete the hybridoma cell strain of anti-B-IgG IgG monoclonal antibody and utilize the monoclonal antibody of its generation to set up immunological detection method.
The invention provides hybridoma cell strain 9C6, this cell strain has been preserved in Chinese Typical Representative culture collection center (CCTCC) on November 28th, 2013, preservation address is, China, Wuhan, Wuhan University, deposit number is CCTCC NO.C2013183, Classification And Nomenclature is mouse hybridoma cell 9C6.
Anti-B-IgG IgG monoclonal antibody, the hybridoma cell strain 9C6 secretion that it is CCTCC NO.C2013183 by deposit number produces.This anti-B-IgG IgG monoclonal antibody can be identified Immunoglobulin IgG1 and IgG2.
The application of anti-B-IgG IgG monoclonal antibody in B-IgG IgG measures.
Beneficial effect of the present invention is:
(1) hybridoma cell strain 9C6 provided by the invention can be for the preparation of the anti-B-IgG IgG of high-titer monoclonal antibody, and tiring that anti-B-IgG IgG mouse ascites antibody ELISA immuning adsorpting analysis (ELISA) method records can reach 5.12 × 10
5(antigen coated concentration 2 μ g/mL).
(2) anti-B-IgG IgG monoclonal antibody provided by the invention is highly sensitive, specificity good, all be less than 1% with the cross reacting rate of bovine casein, milk albumin, Cow Milk ferritin, bovine serum albumin, Protalbinic acid, do not react with goat IgG, sheep IgG, rabbit igg and ox IgM.
(3) anti-B-IgG IgG monoclonal antibody provided by the invention can be applicable to measure the B-IgG IgG content in the sample such as bovine coloctrum and goods, blood.
(4) gold-marking immunity chromatography provided by the invention can be applicable to field quick detection and the quality of production control of the B-IgG IgG content in the sample such as bovine coloctrum and goods, bovine blood.
Brief description of the drawings
Fig. 1 is the front view of the B-IgG IgG immunochromatographydetection detection card prepared of the embodiment of the present invention 3.
Fig. 2 is the sectional elevation of the B-IgG IgG immunochromatographydetection detection card prepared of the embodiment of the present invention 3.
The B-IgG IgG immuno-chromatographic test paper strip that Fig. 3 provides for the application embodiment of the present invention 3 detects the result process decision chart of sample.
Embodiment
Embodiment 1: the screening of hybridoma cell strain 9C6
1. animal immune
Choose 4 of female BALB/c mouse in 6 week age, produced article No. I5506 with B-IgG IgG(by Sigma-Aldrich company) immunity 3 times, first 100 μ g/ only, equivalent complete Freund's adjuvant, fully emulsified, abdominal injection; After 2 weeks, carry out immunity for the second time, 100 μ g/, equivalent incomplete Freund's adjuvant, fully emulsified, abdominal injection; After 2 weeks, carry out immunity for the third time, operation is with immunity for the second time.Measure serum titer, select serum titer soprano at first 3 days booster immunizations of fusion, 100 μ g/ only, do not add adjuvant, abdominal injection.
2. cytogamy
After booster immunization 3 days, get according to a conventional method mouse boosting cell, merge with murine myeloma cell SP2/0.Adopt polyoxyethylene glycol (PEG, molecular weight is 1450) to make fusogen, carry out according to a conventional method the cultivation of fused cell.
3. the screening of cell strain and clone
8-14 days after cytogamy, cell colony grows to the 1/4-1/2 size of hole floorage, and antibody test is carried out in nutrient solution flavescence.Adopt improved sandwich ELISA method to carry out antibody test to the culture hole that has Growth of Hybridoma Cell.Adopt limiting dilution assay to clone, after clone, within about 8-12 days, adopt to use the same method and detect, so after repeated cloning 2-3 time, acquisition hybridoma cell strain 9C6.
Wherein, the concrete steps of improved sandwich ELISA method detection hybridoma supernatant antibody are:
(1) coated: with coating buffer by 1000 times of goat-anti ox IgG antibody (Sigma-Aldrich company, B1645) dilutions.In each reacting hole of enzyme plate, add 100 μ L, 4 DEG C spend the night (or 37 DEG C of incubation 2 ~ 3h).Next day, discards solution in hole, and each hole adds PBS-T lavation buffer solution 220 μ L, vibrates 1 minute, gets rid of liquid in clear opening, then adds PBS-T lavation buffer solution, so washs 3 times.
(2) sealing: add 200 μ L confining liquids in above-mentioned coated reacting hole, put 37 DEG C of incubation 1h.Then wash 3 times.
(3) application of sample: add hybridoma cell strain nutrient solution supernatant 100 μ L in reacting hole, put 37 DEG C of incubation 1h.Then wash 3 times.(do negative control with negative nutrient solution supernatant, do positive control with positive serum, do blank with PBS damping fluid) simultaneously.
(4) add ELIAS secondary antibody: sheep anti mouse enzyme labelled antibody (Sigma-Aldrich company, A3673) the 100 μ L that add new dilution in each reacting hole.37 DEG C of incubation 40min, wash 5 times.
(5) add substrate solution colour developing: in each reacting hole, add the tmb substrate solution 100 μ L that just prepared, 37 DEG C of dark place incubation 10min.
(6) termination reaction: add stop buffer 50 μ L, immediately readings in each reacting hole.
(7) under microplate reader reading: 450nm, read each hole absorbance.
Described coated damping fluid is 1.59g sodium carbonate, 2.93g sodium bicarbonate, and adding distil water, to 1000mL, regulates pH value to 9.6.
Described PBS damping fluid is 8g sodium-chlor, 2.9g disodium hydrogen phosphate, and 0.2g Repone K, 0.2g potassium primary phosphate, adding distil water is settled to 1000mL gained.
Described PBS-T lavation buffer solution is 0.5mL Tween-20, adds PBS damping fluid 1000mL gained.
Described confining liquid is 5.349g ammonium chloride, adding distil water 1000mL gained.
Described substrate solution is A, B liquid TMB nitrite ion (safe day of Jinan and biotechnology company produce).
Described stop buffer is 2M aqueous sulfuric acid.
Embodiment 2: anti-B-IgG IgG monoclonal antibody prepare purifying, hypotype and CHARACTERISTICS IDENTIFICATION
The anti-B-IgG IgG monoclonal antibody hybridoma cell strain 9C6 that embodiment 1 is obtained injects the BALB/c mouse of processing with freund 's incomplete adjuvant in advance, collect the ascites of this mouse, after centrifugal degreasing, adopt Protein A affinity purification method antibody purification (concrete by specification operation).After purifying, immediately to the dialysis of 0.01M pH7.2 phosphate buffered saline buffer, adjust protein concentration to 10mg/mL ,-70 DEG C of refrigerator freezings, for subsequent use;
The hypotype of identifying the anti-B-IgG IgG monoclonal antibody of hybridoma cell strain 9C6 secretion with hypotype identification kit is IgG1.
The tiring of mouse hydroperitoneum antibody of RGDV that records 9C6 with conventional indirect elisa method can reach 5.12 × 10
5(antigen coated concentration 2 μ g/mL), can react with ox IgG1 and IgG2, all be less than 1% with the cross reacting rate of bovine casein, milk albumin, Cow Milk ferritin, bovine serum albumin, Protalbinic acid, do not react with goat IgG, sheep IgG, rabbit igg and ox IgM.
Preparation and the application of embodiment 3 immunochromatographydetection detection cards or test strip
The anti-B-IgG IgG monoclonal antibody that embodiment 2 is obtained is for the preparation of B-IgG IgG immunochromatographydetection detection card, and preparation method comprises the following steps:
(1) preparation of antibody
The anti-B-IgG IgG monoclonal antibody of selecting described cell strain secretion the to produce antibody that serves as a mark, select the anti-ox polyclonal antibody in the sources such as rabbit, ox, horse, donkey as detection line (T line) coated antibody, sheep anti mouse polyclonal antibody is as nature controlling line (C line) coated antibody.
(2) preparation of detection film
T line coated antibody and C line coated antibody are mixed with respectively to the concentration of 0.5-2.0mg/mL with T line coating buffer and C line coating buffer, use the quantitative spray equipment of Bio-dot respectively the two to be sprayed on nitrocellulose filter bar with the interval of 0.4-1cm, discharge rate is 1 μ g/cm, obtain T line and C line, be placed in dry 30-60 minute under 37 DEG C of conditions, then immerse in confining liquid and soak after 10 minutes, dry 3-6 hour under 30 DEG C of conditions, add siccative to seal up for safekeeping, for subsequent use.
Described T line coating buffer is 0.01M pH7.2-7.4 phosphoric acid buffer, and 0.14M sodium-chlor, adds 5g/L sucrose, 0.22 μ m membrane filtration degerming, and 4 DEG C of preservations, were used in 2 weeks.
Described C line coating buffer is 0.01M pH7.2-7.4 phosphoric acid buffer, adds 5g/L sucrose, 0.22 μ m membrane filtration degerming, and 4 DEG C of preservations, were used in 2 weeks.
Described confining liquid is 0.01M pH7.2 PBS damping fluid, adds 10g/L sucrose, 10g/L isinglass, and 0.22 μ m membrane filtration degerming, 4 DEG C of preservations, were used in 1 week.
(3) preparation of sample pad
Glass fibre or polyester sample pad are cut out to growth 300mm, and wide 25mm is rectangular, puts into sample pad pretreatment fluid and soaks after 1 hour, takes out, and under 37 DEG C of conditions, is dried 6 hours, adds siccative to seal up for safekeeping, for subsequent use.The 0.01M pH7.2 phosphoric acid buffer of the tween 20 that described sample pad pretreatment fluid contains 1-10g/L polyvinylpyrrolidone and 0.1-2.0g/L.
(4) preparation of gold mark pad
The preparation of the monoclonal antibody of A, nano gold mark
Before mark by monoclonal antibody containing dialysed overnight in the quadruple distillation water of 0.05g/L sodium-chlor.Get nano-Au solution (grain diameter is 20-40nm), with 0.2 M salt of wormwood adjust pH to 8.2, under the condition of magnetic agitation, by the amount that adds 8-10 μ g antibody in every milliliter of nanometer gold, dropwise add the monoclonal antibody of purifying, stir 15 min.Add oralbumin solution to final concentration 5g/L, continue to stir 10 min.Centrifugal 15 min of 500 × g, remove precipitation, get supernatant, centrifugal 30 min of 12 000 × g.Sucking-off supernatant, the colloidal gold probe of precipitation is also centrifugal with the resuspended liquid redissolution of original volume, and sucking-off supernatant, retains precipitation, adds the gold mark of original volume 1/10 to preserve liquid, and 4 DEG C of preservations are for subsequent use.
Described resuspended liquid is for containing 5g/L oralbumin, and the 0.01M pH7.2 PB damping fluid of 0.2g/L sodium azide, crosses 0.22 μ m filter membrane, 4 DEG C of preservations.
Described mark is preserved liquid for containing 5g/L oralbumin, 100g/L sucrose, and 25g/L trehalose, the 0.01M pH7.2 PB damping fluid of 0.2g/L sodium azide, crosses 0.22 μ m filter membrane, 4 DEG C of preservations.
The preparation of B, gold mark pad
By glass fibre cutting growth 300mm, 5mm width slice, laterally sprays the anti-B-IgG IgG monoclonal antibody of nano gold mark with the discharge rate of 1 μ g/cm then vacuum lyophilization 6h-24h with Bio-dot metal spraying device, add siccative, sealing is preserved.
(5) preparation of absorption pad
Thieving paper is cut out to growth 300mm, and the specification of wide 30mm, obtains absorbent pad;
(6) assembling of test strip: paste successively and detect film, gold mark pad, sample pad and absorbent pad at a corresponding site of PVC backboard, adjacent each several part is overlapping 1mm in junction.
(7) cutting of test strip: cut into wide little of 3mm by assembling test strip with Bio-dot slitting shear machine, obtain B-IgG IgG immuno-chromatographic test paper strip.
(8) assembling of test card, is placed in single part of test strip of well cutting of the present invention in the draw-in groove of plastic bottom card, covers upper cover, compresses, and sees Fig. 1 and Fig. 2.Add siccative, sealing is preserved.
(9) sample buffer formula: 0.05M Tris, 0.3M sodium-chlor, 0.2g/L sodium azide, 0.005M EDTA, pH7.2-7.4.
The application of above-mentioned B-IgG IgG immuno-chromatographic test paper strip:
Sample thief 1mL, then add 24mL sample buffer to mix, getting 200 μ L adds in test card well, leave standstill reading result after 15 minutes, if detection line is aobvious red, nature controlling line line is aobvious red, is judged to positive findings, the content that shows the B-IgG IgG in testing sample is more than or equal to 12mg/mL, sees in Fig. 31; If detection line does not develop the color, nature controlling line line is aobvious red, is judged to negative findings, shows that the content of the B-IgG IgG in testing sample is less than 12mg/mL, sees in Fig. 32; If nature controlling line line does not develop the color, be judged to be invalid detection, see in Fig. 33.
Claims (5)
1. a hybridoma cell strain 9C6, is characterized in that secreting anti-B-IgG IgG monoclonal antibody, and it is preserved in Chinese Typical Representative culture collection center, and deposit number is CCTCC NO.C2013183.
2. an anti-B-IgG IgG monoclonal antibody, hypotype is IgG1, the hybridoma cell strain 9C6 secretion that it is CCTCCNO.C2013183 by deposit number produces, and it is characterized in that the tiring of mouse ascites antibody recording with conventional indirect elisa method can reach 5.12 × 10
5, can react with ox IgG1 and IgG2, be all less than 1% with the cross reacting rate of bovine casein, milk albumin, Cow Milk ferritin, bovine serum albumin, oralbumin, do not react with goat IgG, sheep IgG, rabbit igg and ox IgM.
3. the screening method of the anti-B-IgG IgG monoclonal antibody hybridoma cell of secretion strain, it is characterized in that can detecting in the monoclonal antibody containing having formed immune complex in bovine serum nutrient solution by improved sandwich ELISA pattern, specifically comprise the following steps:
The concrete steps that detect hybridoma supernatant antibody are:
(1) coated: with coating buffer by 1000 times of goat-anti ox IgG antibody dilutions, in each reacting hole of enzyme plate, add 100 μ L, 4 DEG C are spent the night, next day, discards solution in hole, and each hole adds PBS-T lavation buffer solution 220 μ L, vibrate 1 minute, get rid of liquid in clear opening, then add PBS-T lavation buffer solution, so wash 3 times;
(2) sealing: add 200 μ L confining liquids in above-mentioned coated reacting hole, put 37 DEG C of incubation 1h, then wash 3 times;
(3) application of sample: add hybridoma cell strain nutrient solution supernatant 100 μ L in reacting hole, put 37 DEG C of incubation 1h, then wash 3 times, do negative control with negative nutrient solution supernatant, do positive control with positive serum, do blank with PBS damping fluid simultaneously;
(4) add ELIAS secondary antibody: the sheep anti mouse enzyme labelled antibody 100 μ L that add new dilution in each reacting hole.37 DEG C of incubation 40min, wash 5 times;
(5) add substrate solution colour developing: in each reacting hole, add the tmb substrate solution 100 μ L that just prepared, 37 DEG C of dark place incubation 10min;
(6) termination reaction: add stop buffer 50 μ L, immediately readings in each reacting hole;
(7) under microplate reader reading: 450nm, read each hole absorbance.
4. the test card of B-IgG IgG or a preparation for test strip, is characterized in that: comprise the following steps:
(1) preparation of antibody
The anti-B-IgG IgG monoclonal antibody of selecting hybridoma cell strain 9C6 claimed in claim 2 secretion the to produce antibody that serves as a mark, select the anti-ox polyclonal antibody in the sources such as rabbit, ox, horse, donkey as detection line coated antibody, sheep anti mouse polyclonal antibody is as nature controlling line coated antibody;
(2) preparation of detection film
Detection line coated antibody and nature controlling line coated antibody are mixed with respectively to the concentration of 0.5-2.0mg/mL with detection line coating buffer and nature controlling line coating buffer, use the quantitative spray equipment of Bio-dot respectively the two to be sprayed on nitrocellulose filter bar with the interval of 0.4-1cm, discharge rate is 1 μ g/cm, obtain detection line and nature controlling line, be placed in dry 30-60 minute under 37 DEG C of conditions, then immerse in confining liquid and soak after 10 minutes, dry 3-6 hour under 30 DEG C of conditions, add siccative to seal up for safekeeping, for subsequent use;
(3) preparation of sample pad
Glass fibre or polyester sample pad are cut out to growth 300mm, and wide 25mm is rectangular, puts into sample pad pretreatment fluid and soaks after 1 hour, takes out, and under 37 DEG C of conditions, is dried 6 hours, adds siccative to seal up for safekeeping, for subsequent use;
(4) preparation of gold mark pad
The anti-B-IgG IgG monoclonal antibody of nano gold mark is sprayed on glass fibre with the discharge rate of 1 μ g/cm with Bio-dot metal spraying device, then vacuum lyophilization 6h-24h, adds siccative, and sealing is preserved;
(5) preparation of absorption pad
Thieving paper is cut out to growth 300mm, and the specification of wide 30mm, obtains absorbent pad;
(6) assembling of test strip: paste successively and detect film, gold mark pad, sample pad and absorbent pad at a corresponding site of PVC backboard, adjacent each several part is overlapping 1mm in junction;
(7) cutting of test strip: cut into wide little of 3mm by assembling test strip with Bio-dot slitting shear machine;
(8) assembling of test card, is placed in single part of test strip of well cutting of the present invention in the draw-in groove of plastic bottom card, covers upper cover, compresses.Add siccative, sealing is preserved;
(9) sample buffer formula: 0.05M Tris, 0.3M sodium-chlor, 0.2g/L sodium azide, 0.005M EDTA, pH7.2-7.4.
5. the test card of B-IgG IgG according to claim 4 or the preparation of test strip, is characterized in that: the preparation of the gold mark pad of (4) step comprises the following steps:
Before mark by monoclonal antibody containing dialysed overnight in the quadruple distillation water of 0.05g/L sodium-chlor; Getting grain diameter is 20-40nm nano-Au solution, with 0.2 M salt of wormwood adjust pH to 8.2, under the condition of magnetic agitation, by the amount that adds 8-10 μ g antibody in every milliliter of nanometer gold, dropwise adds the monoclonal antibody of purifying, stirs 15 min.Add oralbumin solution to final concentration 5g/L, continue to stir 10 min.Centrifugal 15 min of 500 × g, remove precipitation, get supernatant, centrifugal 30 min of 12 000 × g, sucking-off supernatant, the colloidal gold probe of precipitation is also centrifugal with the resuspended liquid redissolution of original volume, and sucking-off supernatant, retains precipitation, add the gold mark of nano-Au solution original volume 1/10 to preserve liquid, 4 DEG C of preservations, for subsequent use;
Described resuspended liquid is for containing 5g/L oralbumin, and the 0.01M pH7.2 PB damping fluid of 0.2g/L sodium azide, crosses 0.22 μ m filter membrane, 4 DEG C of preservations;
Described mark is preserved liquid for containing 5g/L oralbumin, 100g/L sucrose, and 25g/L trehalose, the 0.01M pH7.2 PB damping fluid of 0.2g/L sodium azide, crosses 0.22 μ m filter membrane, 4 DEG C of preservations.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410247248.8A CN104099300B (en) | 2013-11-22 | 2014-06-06 | Hybridoma cell strain and its application of anti-B-IgG IgG monoclonal antibody can be secreted |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310595381.8 | 2013-11-22 | ||
| CN201310595381 | 2013-11-22 | ||
| CN2013105953818 | 2013-11-22 | ||
| CN201410247248.8A CN104099300B (en) | 2013-11-22 | 2014-06-06 | Hybridoma cell strain and its application of anti-B-IgG IgG monoclonal antibody can be secreted |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104099300A true CN104099300A (en) | 2014-10-15 |
| CN104099300B CN104099300B (en) | 2017-03-08 |
Family
ID=51667842
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410247248.8A Expired - Fee Related CN104099300B (en) | 2013-11-22 | 2014-06-06 | Hybridoma cell strain and its application of anti-B-IgG IgG monoclonal antibody can be secreted |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104099300B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785013A (en) * | 2016-04-21 | 2016-07-20 | 卢连伟 | Colloidal gold immunochromatographic test strip for aided detection of pancreatic cancer and preparation method of golloidal gold immunochromatographic test strip |
| CN107389940A (en) * | 2017-08-23 | 2017-11-24 | 潍坊市康华生物技术有限公司 | A kind of vaginitis multi-link detection reagent kit and preparation method thereof |
| CN107748263A (en) * | 2017-08-31 | 2018-03-02 | 北京臻惠康生物科技有限公司 | The new application and kit of a kind of plasminogen |
| CN109507421A (en) * | 2018-12-07 | 2019-03-22 | 广西大学 | Water resistant ox IgG monoclonal antibody cell strain and its preparation method and application |
| CN109507420A (en) * | 2018-12-07 | 2019-03-22 | 广西大学 | The rapid detection method of buffalo Fasciola gigantica antibody |
| CN111896750A (en) * | 2020-08-07 | 2020-11-06 | 杭州都林生物科技有限公司 | Bovine immunoglobulin (IgG) double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method |
| CN114264819A (en) * | 2021-12-08 | 2022-04-01 | 上海理工大学 | Quantum dot nanosphere immunochromatography test strip for rapidly detecting new coronavirus |
| CN114805589A (en) * | 2022-06-14 | 2022-07-29 | 郑州伊美诺生物技术有限公司 | Monoclonal antibody capable of simultaneously recognizing antibodies of cattle, goats and sheep |
| CN114894911A (en) * | 2022-03-18 | 2022-08-12 | 辽宁成大生物股份有限公司 | Method for controlling bovine serum product quality |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103197077A (en) * | 2013-03-20 | 2013-07-10 | 郑州伊美诺生物技术有限公司 | Assay kit for detecting trace bovine immunoglobulin G |
-
2014
- 2014-06-06 CN CN201410247248.8A patent/CN104099300B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103197077A (en) * | 2013-03-20 | 2013-07-10 | 郑州伊美诺生物技术有限公司 | Assay kit for detecting trace bovine immunoglobulin G |
Non-Patent Citations (3)
| Title |
|---|
| 李忠秋 等: "免疫胶体金半定量检测牛初乳IgG含量方法的建立及初步应用", 《中国奶牛》, no. 1, 31 December 2006 (2006-12-31) * |
| 杨鼎 等: "胶体金免疫层析技术研究进展", 《畜牧与饲料科学》, vol. 34, no. 12, 31 December 2013 (2013-12-31) * |
| 韩秀娥 等: "牛初乳免疫球蛋白IgG单克隆抗体的制备", 《黑龙江畜牧兽医》, no. 7, 31 December 2011 (2011-12-31) * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785013A (en) * | 2016-04-21 | 2016-07-20 | 卢连伟 | Colloidal gold immunochromatographic test strip for aided detection of pancreatic cancer and preparation method of golloidal gold immunochromatographic test strip |
| CN107389940A (en) * | 2017-08-23 | 2017-11-24 | 潍坊市康华生物技术有限公司 | A kind of vaginitis multi-link detection reagent kit and preparation method thereof |
| CN107748263A (en) * | 2017-08-31 | 2018-03-02 | 北京臻惠康生物科技有限公司 | The new application and kit of a kind of plasminogen |
| CN109507421B (en) * | 2018-12-07 | 2022-02-01 | 广西大学 | Anti-buffalo IgG monoclonal antibody cell strain and preparation method and application thereof |
| CN109507420A (en) * | 2018-12-07 | 2019-03-22 | 广西大学 | The rapid detection method of buffalo Fasciola gigantica antibody |
| CN109507421A (en) * | 2018-12-07 | 2019-03-22 | 广西大学 | Water resistant ox IgG monoclonal antibody cell strain and its preparation method and application |
| CN109507420B (en) * | 2018-12-07 | 2022-02-01 | 广西大学 | Rapid detection method of buffalo fasciola gigantica antibody |
| CN111896750A (en) * | 2020-08-07 | 2020-11-06 | 杭州都林生物科技有限公司 | Bovine immunoglobulin (IgG) double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method |
| CN114264819A (en) * | 2021-12-08 | 2022-04-01 | 上海理工大学 | Quantum dot nanosphere immunochromatography test strip for rapidly detecting new coronavirus |
| CN114894911A (en) * | 2022-03-18 | 2022-08-12 | 辽宁成大生物股份有限公司 | Method for controlling bovine serum product quality |
| CN114894911B (en) * | 2022-03-18 | 2023-10-24 | 辽宁成大生物股份有限公司 | Method for controlling quality of bovine serum products |
| CN114805589A (en) * | 2022-06-14 | 2022-07-29 | 郑州伊美诺生物技术有限公司 | Monoclonal antibody capable of simultaneously recognizing antibodies of cattle, goats and sheep |
| CN114805589B (en) * | 2022-06-14 | 2024-03-26 | 郑州伊美诺生物技术有限公司 | Monoclonal antibody capable of simultaneously recognizing cow, goat and sheep antibodies |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104099300B (en) | 2017-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104099300A (en) | Hybridoma cell line able to secrete anti-bovine immunoglobulin IgG monoclonal antibody and application thereof | |
| CN101993855B (en) | Hybridoma cell line 1C11 and anti-aflatoxin general monoclonal antibody generated by same as well as applications thereof | |
| CN103173420B (en) | Hybridoma cell capable of secreting anti-cardiac troponin I monoclonal antibodies and applications thereof | |
| CN110068688B (en) | Lactoferrin competition method nanoflower immunity detection flow chromatography detection card in cow's milk | |
| RU2005106230A (en) | SPECIFIC ANTIBODIES FOR THE DIAGNOSIS OF HEART FAILURE | |
| CN102675463A (en) | Carbendazim monoclonal antibody, preparation method and application thereof | |
| CN102879561B (en) | Screening test paper strip for human lactoferrin in milk and preparation method | |
| CN102876634B (en) | PMSG (pregnant mare serum gonadotropin) double-antibody sandwich ELISA (enzyme-linked immunosorbent assay) kit | |
| CN108700576A (en) | The system for preparing the application of the method, kit and kit of antibody pair and preparing antibody pair | |
| CN101307303B (en) | Kit for detecting clenobuterol hydrochloride residue and method for preparing same | |
| CN103333864B (en) | Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody | |
| CN102236015A (en) | Competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors and preparation method of competitive inhibiting enzyme-linked immune chip kit | |
| CN111537739A (en) | Monoclonal antibody titer rapid determination immunofluorescence test strip | |
| CN103215230A (en) | Hybridoma cell strain AFM1B7, monoclonal antibody thereof and aflatoxin M1 flow lag immunization time-resolved fluorescence quick test kit | |
| CN107271685A (en) | A kind of method and its dedicated kit for detecting the content of fowl leukocyte interleukin 2 | |
| CN109596839A (en) | People and peptide element fast quantitative measurement method for detecting and kit | |
| CN108918851A (en) | A kind of preparation method of Lamotrigine colloidal gold strip | |
| CN107389920B (en) | A kind of method and its dedicated kit detecting fowl leukocyte interleukin 17a content | |
| CN102590517A (en) | Immunochromatography test paper and preparation method thereof | |
| CN105732810A (en) | Procalcitonin monoclonal antibody and application thereof | |
| CN104749371B (en) | People's nephroblastoma overepressed gene encoding proteins enzyme linked immunological kit | |
| CN113025580B (en) | Hybridoma cell strain, anti-fipronil monoclonal antibody produced by hybridoma cell strain and application of anti-fipronil monoclonal antibody | |
| CN111748528B (en) | Hybridoma cell strain secreting monoclonal antibody against fipronil and metabolite thereof and application of hybridoma cell strain | |
| CN104004718A (en) | Universal monoclonal antibody hybridoma cell strain capable of resisting pirlimycin and application thereof | |
| CN107860911A (en) | A kind of preparation method of pathogenic salmonella test-strips |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170308 Termination date: 20170606 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |