CN102942531B - The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody - Google Patents
The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody Download PDFInfo
- Publication number
- CN102942531B CN102942531B CN201210438821.4A CN201210438821A CN102942531B CN 102942531 B CN102942531 B CN 102942531B CN 201210438821 A CN201210438821 A CN 201210438821A CN 102942531 B CN102942531 B CN 102942531B
- Authority
- CN
- China
- Prior art keywords
- trimeric cyanamide
- melamine
- preparation
- melamine derivative
- monoclonal antibody
- 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.)
- Active
Links
- GVRROEPNWSIRQU-UHFFFAOYSA-N CN1C(NC(CCCC(O)=O)=O)N=C(N)NC1N Chemical compound CN1C(NC(CCCC(O)=O)=O)N=C(N)NC1N GVRROEPNWSIRQU-UHFFFAOYSA-N 0.000 description 1
- 0 NC1=NC(N)=NC(N)=*C1 Chemical compound NC1=NC(N)=NC(N)=*C1 0.000 description 1
- FPNJOSSDGMOPHF-UHFFFAOYSA-N Nc1nc(N)nc(NC(CCCC(O)=O)=O)n1 Chemical compound Nc1nc(N)nc(NC(CCCC(O)=O)=O)n1 FPNJOSSDGMOPHF-UHFFFAOYSA-N 0.000 description 1
- VANNPISTIUFMLH-UHFFFAOYSA-N O=C(CCC1)OC1=O Chemical compound O=C(CCC1)OC1=O VANNPISTIUFMLH-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of trimeric cyanamide to spread out the preparation method and application of (Melamine) biological and trimeric cyanamide monoclonal antibody, the preparation method of described melamine derivative comprises the following steps: a) by melamine goes in the dimethylformamide being preheated to preset temperature, add Pyroglutaric acid and reacted for first scheduled time; B) add ether by after reaction after solution cooling, produce precipitation; C) filter described precipitation, and solid after filtration is carried out recrystallization, obtain melamine derivative.According to the melamine derivative of the embodiment of the present invention, an arm structure has been had more relative to trimeric cyanamide, can be connected with carrier proteins by this arm structure, connect gained antigen immunogenicity good, high specificity, specific antibody can be produced by stimulating animal, and height of tiring, solve and be difficult to the problem that independent melamine compound prepares potent antibodies.<!-- 0001 -->
Description
Technical field
The present invention relates to chemosynthesis technical field, more specifically, the present invention relates to the preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody.
Background technology
Melamine compound because of molecule little, structure is simple, after direct coupling carrier albumen, micromolecular space structure can not be exposed fully, be unfavorable for this micromolecular compound of lymphocyte identification, so the antibody titer prepared with this antigen of melamine molecule direct coupling carrier albumen without transformation is low, poor specificity, cannot meet the demand of the research and development of detection kit.
Summary of the invention
The present invention is intended at least one of solve the problems of the technologies described above.
For this reason, one object of the present invention is to propose a kind ofly to be connected with carrier proteins that afterwards antibody titer is high, the melamine derivative of high specificity.
According to the melamine derivative of the embodiment of the present invention, there is following molecular structural formula:
According to the melamine derivative of the embodiment of the present invention, an arm structure has been had more relative to trimeric cyanamide, can be connected with carrier proteins by this arm structure, connect gained antigen immunogenicity good, high specificity, specific antibody can be produced by stimulating animal, and height of tiring, solve and be difficult to the problem that independent melamine compound prepares potent antibodies.
The invention allows for the preparation method of melamine derivative, it specifically can comprise the following steps:
A) trimeric cyanamide (1.26g, 10mmol) is dissolved in is preheated in the dimethylformamide (DMF40mL) of 40 DEG C ~ 50 DEG C, add Pyroglutaric acid (1.14g, 10mmol) and react 2h;
B) add ether 10mL by after reaction after solution cooling, produce white precipitate;
C) filter described precipitation, and solid after filtration is carried out recrystallization, 1.8g melamine derivative can be obtained.
Operation is simple for the method, and preparation cost is low, and it is of many uses to prepare gained melamine derivative, is applicable to promoting the use of.
Another object of the present invention is that proposing a kind of melamine derivative is preparing the application in trimeric cyanamide monoclonal antibody.
An arm structure has been had more relative to trimeric cyanamide based on described melamine derivative, can be connected with carrier proteins by this arm structure, connect gained antigen immunogenicity good, high specificity, therefore, can by described trimeric cyanamide for the preparation of trimeric cyanamide monoclonal antibody.
Present invention also offers the preparation method of trimeric cyanamide monoclonal antibody, its concrete operation step can comprise:
A) by melamine derivative and hemocyanin (KLH) by coupling agent coupling, obtain complete antigen;
B) by melamine derivative and bovine serum albumin (BSA) by coupling agent coupling, obtain trimeric cyanamide-protein conjugate (trimeric cyanamide-BSA);
C) by immunity test animal after described complete antigen and Freund's complete adjuvant or Freund's incomplete adjuvant emulsification, and the tenth day extracting vein blood after the 4th immunity measures antibody titer;
D) by the splenocyte of experimental animal and myeloma cell fusion, separation screening cell strain of monoclonal antibody;
E) by the stable positive clone strain enlarged culturing of screening, inoculation test animal, preparation monoclonal antibody ascites;
F) described antibody ascites is separated supernatant, with ProteinA affinity chromatography column separating purification, obtains trimeric cyanamide monoclonal antibody.
The complete antigen preparing gained according to aforesaid method may be used for the immunity of animal, and the trimeric cyanamide-BSA obtained may be used for detecting antibody.
In an embodiment of the present invention, about the method measuring antibody titer, be not particularly limited, such as, can adopt competitive ELISA method, its concrete operations can be:
Wrap by elisa plate with trimeric cyanamide-BSA, close with 1%BSA, detersive enzyme yoke plate, mice serum is diluted to different ratios, and with the antigen-reactive on elisa plate, before immunity, mice serum does negative control, detersive enzyme yoke plate, then answer with two anti-reflective of horseradish peroxidase-labeled, detersive enzyme yoke plate, hydrogen peroxide is as substrate, TMB (3,3 ', 5,5 '-tetramethyl benzidine) colour developing, 2NH
2sO
4as stop buffer, measure OD value in 450nm.Wherein, detersive enzyme yoke plate washings used is 0.01MPBS, is the tween 20 of 0.05% containing volume basis hundred, and pH value is 7.4.
Another object of the present invention is to propose the application of a kind of melamine derivative in the test kit for the preparation of detection trimeric cyanamide.By the application of this test kit, the residual quantity of the Ciprofloxacin in analyzing animal urine, blood, muscle, liver, milk and feed equal samples can be detected, also can be used for the diagnosis and prognosis of cancer, cardiovascular diseases, diabetes or some other Diseases, there is the advantages such as accurate, special, sensitive, stable, convenient.
Present invention also offers a kind of elisa kit for detecting detecting human vascular endothelial growth factor (VEGF); comprise in kit: the enzyme conjugates that the antibody marked from the enzyme-linked reaction plate of the vegf receptor bag quilt of insect cell expression purifying, horseradish peroxidase (HRP) configures with protective material, as the lyophilized powder of the vegf protein of positive control, as negative control protein lyophilized powder, sample diluting liquid, PBST concentrated bleaching liquid, 3; 3 '; 5, the chromogenic substrate of 5 '-tetramethyl benzidine configuration, the 2NH as stop buffer
2sO
4, shrouding gummed paper.This test kit can be used for the diagnosis and prognosis of cancer, cardiovascular diseases, diabetes or some other Diseases, has the advantages such as accurate, special, sensitive, stable, convenient.Meanwhile, present invention also offers the method using insect cell high expression foreign protein.
Additional aspect of the present invention and advantage will part provide in the following description, and part will become obvious from the following description, or be recognized by practice of the present invention.
Accompanying drawing explanation
Above-mentioned and/or additional aspect of the present invention and advantage will become obvious and easy understand from accompanying drawing below combining to the description of embodiment, wherein:
Fig. 1 is the canonical plotting of the OD450 mean value according to one embodiment of the invention Plays product.
Embodiment
Further describe the present invention below by way of specific embodiment, the following example for illustration of object, but not for limiting the scope of the invention.The test method of unreceipted actual conditions in the following example, can operate according to the condition described in common molecular cloning handbook.Test materials used in the following example, if no special instructions, is and buys gained from routine biochemistry reagent shop.
The preparation of embodiment 1 melamine derivative
One, test materials
Trimeric cyanamide, DMF, Pyroglutaric acid, ether are buys gained from routine biochemistry reagent shop.
Two, testing sequence
A) trimeric cyanamide (1.26g, 10mmol) is dissolved in the DMF(40mL being preheated to 40 DEG C ~ 50 DEG C) in, add Pyroglutaric acid (1.14g, 10mmol) and react 2h;
B) add ether 10mL by after reaction after solution cooling, produce white precipitate;
C) filter described precipitation, and solid after filtration is carried out recrystallization, 1.8g melamine derivative can be obtained.
The preparation of embodiment 2 trimeric cyanamide monoclonal antibody
One, test materials
Melamine derivative (preparing gained by embodiment 1);
KLH, BSA, carbodiimide, Freund's complete adjuvant or Freund's incomplete adjuvant, hydrogen peroxide, TMB, H
2sO
4for buying gained from routine biochemistry reagent shop;
Washings is 0.01MPBS, is the tween 20 of 0.05% containing volume basis hundred, and pH value is 7.4, can allocate gained voluntarily.
Two, experimental animal
Balb/c mouse (Test Animal Centre, Academy of Military Medical Sciences, P.L.A provides)
Three, testing sequence
A) melamine derivative and KLH are passed through carbodiimide coupling, obtain complete antigen;
B) melamine derivative and BSA are passed through carbodiimide coupling, obtain trimeric cyanamide-protein conjugate (trimeric cyanamide-BSA), detection antigen;
C) by Balb/c mouse immune after described complete antigen and Freund's complete adjuvant or Freund's incomplete adjuvant emulsification, and the tenth day extracting vein blood after the 4th immunity measures antibody titer;
D) by the splenocyte of Balb/c mouse and myeloma cell fusion, separation screening cell strain of monoclonal antibody;
E) by the stable positive clone strain enlarged culturing of screening, inoculation Balb/c mouse, preparation monoclonal antibody ascites;
F) described antibody ascites is separated supernatant, with ProteinA affinity chromatography column separating purification, obtains trimeric cyanamide monoclonal antibody.
Wherein, about the method measuring antibody titer, be not particularly limited, such as, can adopt competitive ELISA method, its measuring principle is:
Specific binding according to antigen-antibody reacts, and adopts competitive ELISA method, enzyme plate capillary strip wraps by anti-melamine mouse monoclonal antibody IgG, adds the standard substance of trimeric cyanamide or sample extract and trimeric cyanamide-HRP cross-linking agent.The binding site of the melamine molecule competition binding trimeric cyanamide specific antibody that free melamine molecule and HRP are cross-linked.Combined melamine molecule and HRP-trimeric cyanamide is not had to be washed removing, develop the color with tmb substrate, blueness is become under horseradish peroxidase effect, after adding stop buffer, color becomes yellow from blueness, light absorption value is measured at 450nm place, sample light absorption value becomes negative correlation with the content of its contained residual melamine molecule, is multiplied by its corresponding dilution factor more again, can draws the residual content of trimeric cyanamide in sample with typical curve.
Embodiment 3 is for detecting the detection experiment of the test kit of trimeric cyanamide
One, test materials
1,96 hole microwell plates (wrapping by trimeric cyanamide monoclonal antibody) 1 plate (12 × 8 hole)
2, trimeric cyanamide standard substance liquid: 6 pipes (0.5ml/ pipe), are labeled as standard substance 1-6 respectively.Sample should be deposited in the cool, or is placed in 4 DEG C of refrigerators and stores.
Standard substance 1:1 manages (content of melamine is 0ng/ml);
Standard substance 2:1 manages (content of melamine is 0.1ng/ml);
Standard substance 3:1 manages (content of melamine is 0.5ng/ml);
Standard substance 4:1 manages (content of melamine is 1ng/ml);
Standard substance 5:1 manages (content of melamine is 5ng/ml);
Standard substance 6:1 manages (content of melamine is 10ng/ml).
3, enzyme connection thing (HRP-Melamine concentrated solution): 1 pipe (50ul).During use, 1:1000 doubly dilutes, now with the current.
4, developer A(white): 1 bottle (8ml).
5, developer B(black): 1 bottle (8ml).
6, reaction terminating liquid (brown): 1 bottle (15ml).
7, diluted sample/wash plate concentrated solution: 1 bottle (40ml), dilutes with distilled water 1:25 during use.
8, fresh pork, pig liver, Ren sus domestica, egg, milk, milk powder.
Two, testing installation
Microwell plate enzyme mark detector;
Refrigerated centrifuge;
Micropipet: 0.5-10 μ l, 20-200 μ l and 200-1000 μ l;
Suction nozzle: 0.5-10 μ l, 20-200 μ l and 200-1000 μ l.
Three, testing sequence
1, preparation of reagents
By diluted sample/washing plate concentrated solution distilled water 1:25 doubly dilutes, obtain diluting working fluid;
Use dilution working fluid, enzyme is joined thing and dilutes with 1:1000, obtain enzyme connection thing working fluid.
2, sample process
1) fresh pork, pig liver, Ren sus domestica are shredded homogenate, the unnecessary homogenate of liquid milk product such as egg, milk, milk powder (can add water);
2) add 4ml70% methyl alcohol in the sample that 1g homogenate is good or 1ml liquid milk product and acutely mix 10min(jerk up and down or vortex is mixed);
3), at 4 DEG C, the centrifugal 10min of 3000rpm, gets supernatant;
4) dilution working fluid 1:2 is utilized to dilute supernatant;
5) with 50ul/ hole, add in microwell plate for detecting.
3, detecting step
1) before experiment, all kit components are balanced 30 minutes at room temperature;
2) prepared and diluted/wash plate working fluid (see table 1);
Table 1 dilutes/washes plate working fluid preparation permutation table
3) trimeric cyanamide-HRP cross-linking agent working fluid is prepared;
4) take plank apart, in microwell plate support, insert the micropore lath tested for standard substance and sample detection of sufficient amount, the position of record standard product and sample;
5) the standard substance liquid or the ready detected sample that add 50 μ l/ holes also arrange repeating hole simultaneously.New rifle head should be used when adding each new standard substance or sample;
6) in each hole, the enzyme-Melamine cross-linking agent working fluid that 50 μ l have diluted is added, in table 2;
The each hole of table 2 adds scale
| Standard substance | Sample extract | Inner quality control liquid | Enzyme-CIP working fluid | |
| Standard sample wells | 50ul | / | / | 50ul |
| Sample well | / | 50ul | / | 50ul |
| Inner quality control hole | / | / | 50ul | 50ul |
7) shake plank, make it mix, and be placed on 37 DEG C of thermostat containers and hatch 30min;
8) poured out by the liquid in hole and pat dry on thieving paper, wash plate with diluting/washing plate working fluid, every hole adds 200-250 μ l, and washing pats dry, and repeats 3-5 time;
9) substrate/developer (A:B=1:1) adding 100 μ l in every hole slightly shakes mixing, hatches 5 minutes in 37 ° of C thermostat container lucifuges;
10) in every hole, add 100 μ l stop buffers, slight concussion mixing also surveys light absorption value at 450nm place, detects after adding stop buffer within 30min.
Four, test-results
While detection sample, standard substance should be used to do parallel laboratory test.With log(100 × standard concentration) be ordinate zou, OD
450be worth for X-coordinate draws linear standard curve.According to the OD that sample measures
450value and dilution factor, calculate mensuration content ng/ml(and ppb of sample trimeric cyanamide, with 1ml liquid for 1g calculates).
Content according to the trimeric cyanamide in following formulae discovery sample:
Content=analytical concentration × dilution factor the ppb of trimeric cyanamide
The dilution factor of various sample is in table 3.
The dilution factor of each sample of table 3
| Sample | Dilution factor |
| Pork | 20 |
| Liver | 20 |
| Kidney | 20 |
| Egg | 20 |
| Milk | 20 |
The mean value that 1.1 standard substance measure
The mean value that trimeric cyanamide standard substance liquid standard substance 1-6 measures is as shown in table 4.
The mean value of table 4 standard substance 1-6
| Standard substance | Standard substance final concentration | OD450 mean value (n=10) |
| Standard substance 6 | 10ppb | 0.099 |
| Standard substance 5 | 5ppb | 0.157 |
| Standard substance 4 | 1ppb | 0.427 |
| Standard substance 3 | 0.5ppb | 0.583 |
| Standard substance 2 | 0.1ppb | 0.837 |
| Standard substance 1 | 0ppb | 1.219 |
1.2 mean values measured according to standard substance, drawing standard curve also does linear regression, and described typical curve as shown in Figure 1.
1.3 calculate the analytical concentration of trimeric cyanamide according to regression formula.
Analytical concentration (ng/ml)=1/100 × 10^(-2.572 × mensuration OD value+3.17 of trimeric cyanamide)
The analytical concentration of trimeric cyanamide corresponding under different inhibiting rate is in table 5.
Trimeric cyanamide analytical concentration table corresponding under the different inhibiting rate of table 5
| Competitive assays rate (%) | OD 450 | Analytical concentration ng/ml |
| 10 | 1.097 | 0.02 |
| 20 | 0.975 | 0.05 |
| 30 | 0.853 | 0.09 |
| 40 | 0.731 | 0.19 |
| 50 | 0.610 | 0.40 |
| 60 | 0.488 | 0.82 |
| 70 | 0.366 | 1.70 |
| 80 | 0.244 | 3.49 |
| 90 | 0.122 | 7.19 |
Known according to table 5: IC50 is 0.4ng/ml.
Suppress to suppress interval computation for determining more than 20% according to inhibiting rate, Determination Limit is 0.05ng/ml, linearity range 0.05 ~ 10ng/ml.
The concentration of trimeric cyanamide in 1.4 samples
In sample, the concentration of trimeric cyanamide is in table 6.
The concentration of trimeric cyanamide in table 6 sample
| Sample | Concentration ppb |
| Pork | Analytical concentration × 20 |
| Liver | Analytical concentration × 20 |
| Kidney | Analytical concentration × 20 |
| Egg | Analytical concentration × 20 |
| Milk | Analytical concentration × 20 |
Can find out according to above-described embodiment, simple according to the melamine derivative preparation method of the embodiment of the present invention, prepare gained trimeric cyanamide monoclonal antibody identifiable design melamine molecule according to described melamine derivative, high specificity, highly sensitive, good stability; Also can be applicable to prepare the test kit detecting trimeric cyanamide, detection method simple possible, and Detection results is good, opens new approaches for developing other micromolecular compound detection kit simultaneously.
Although illustrate and describe embodiments of the invention, those having ordinary skill in the art will appreciate that: can carry out multiple change, amendment, replacement and modification to these embodiments when not departing from principle of the present invention and aim, scope of the present invention is by claim and equivalents thereof.
Claims (1)
1. a preparation method for melamine derivative, is characterized in that, comprises the following steps:
A) by melamine goes in the dimethylformamide being preheated to preset temperature, add Pyroglutaric acid and reacted for first scheduled time, described preset temperature is 40 DEG C ~ 50 DEG C, and described first scheduled time is 2h;
B) add ether by after reaction after solution cooling, produce precipitation;
C) filter described precipitation, and solid after filtration is carried out recrystallization, obtain melamine derivative, its molecular structural formula is:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210438821.4A CN102942531B (en) | 2012-11-06 | 2012-11-06 | The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210438821.4A CN102942531B (en) | 2012-11-06 | 2012-11-06 | The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102942531A CN102942531A (en) | 2013-02-27 |
| CN102942531B true CN102942531B (en) | 2015-12-16 |
Family
ID=47725547
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210438821.4A Active CN102942531B (en) | 2012-11-06 | 2012-11-06 | The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102942531B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110938040B (en) * | 2019-11-25 | 2021-03-09 | 广东达元绿洲食品安全科技股份有限公司 | Melamine hapten and artificial antibody as well as preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102086228A (en) * | 2009-12-04 | 2011-06-08 | 北京维德维康生物技术有限公司 | ELISA (Enzyme Linked Immunosorbent Assay) Kit and application thereof |
-
2012
- 2012-11-06 CN CN201210438821.4A patent/CN102942531B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102086228A (en) * | 2009-12-04 | 2011-06-08 | 北京维德维康生物技术有限公司 | ELISA (Enzyme Linked Immunosorbent Assay) Kit and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 三聚氰胺单克隆抗体研制及应用研究;陈昕;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20110515(第05期);22、25、30、32－33、41 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102942531A (en) | 2013-02-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2007257105B2 (en) | Elisa kit for detecting Sudan red and method thereof | |
| CN103575886B (en) | The enzyme linked immunological kit of detection ochratoxin A and application thereof | |
| CN103869070B (en) | Detect enzyme linked immunological kit and the application thereof of dibutyl phthalate | |
| CN104897864B (en) | The enzyme linked immunological kit of detection amantadine and application thereof | |
| CN101776685B (en) | Enzyme linked immunosorbent assay kit for detecting trimethoprim medicament and application thereof | |
| CN103288872B (en) | Parathion-methyl hapten and its preparation method and application | |
| CN102206270B (en) | Saxitoxin artificial antigen and anti-saxitoxin antibody, their preparation methods and application | |
| CN104076154A (en) | Enzyme linked immunosorbent assay kit detecting folic acid and application thereof | |
| CN103421072A (en) | Dexamethasone semi-antigen, preparation method and applications thereof | |
| CN109180519B (en) | Olaquindox metabolite antigen, antibody, enzyme-linked immunosorbent assay kit and detection method | |
| CN105319368A (en) | Enzyme linked immunosorbent assay kit used for detecting zearalenone, and detection method thereof | |
| CN102942531B (en) | The preparation method and application of a kind of melamine derivative and trimeric cyanamide monoclonal antibody | |
| CN105758846A (en) | Chemiluminescence enzyme-linked immunosorbent assay reagent kit for detecting clenbuterol | |
| CN102331500A (en) | Method and enzyme linked immunosorbent assay kit for detecting lemon yellow | |
| CN103145633B (en) | Novel melamine antigen and antibody and application | |
| CN116008524A (en) | Protein-free universal protection liquid, preparation method thereof and application thereof in mycotoxin quantitative detection kit | |
| CN104730250A (en) | Enzyme linked immunosorbent assay kit for detecting human kidney injury molecule-1 | |
| CN105503757A (en) | Preparation methods of melamine hapten and melamine antigen, and application thereof in quantum dot fluorescence immunoassay kit | |
| CN107228939A (en) | Detect enzyme linked immunological kit and its application of morphine and codeine | |
| CN104569398B (en) | The enzyme linked immunological kit of detection ethoxy quinoline and application thereof | |
| CN103420879B (en) | Dapsone hapten and its preparation method and application | |
| CA2835068C (en) | Method for immunologically measuring soluble lr11 | |
| CN105974127A (en) | Human neutrophil apolipoprotein heterodimer quantifying device based on enzyme-linked immune adsorption technology | |
| CN112142756A (en) | Preparation of aflatoxin M1 hapten and holoantigen as well as preparation and application of rapid transfer quantitative kit | |
| CN105503759A (en) | Preparation methods of melamine hapten and melamine antigen, and application thereof in chemiluminescent immunoassay kit |
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 |