CN102382189B - Synthesis method of 5-hydroxymethylfurfural complete antigen - Google Patents
Synthesis method of 5-hydroxymethylfurfural complete antigen Download PDFInfo
- Publication number
- CN102382189B CN102382189B CN201110228269.1A CN201110228269A CN102382189B CN 102382189 B CN102382189 B CN 102382189B CN 201110228269 A CN201110228269 A CN 201110228269A CN 102382189 B CN102382189 B CN 102382189B
- Authority
- CN
- China
- Prior art keywords
- hydroxymethylfurfural
- liquid
- serum albumin
- bovine serum
- hydroxymethyl furfural
- 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
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a synthesis method of a 5-hydroxymethylfurfural complete antigen, which dissolves 5-hydroxymethylfurfural into a glutaraldehyde solution, so a mixed liquor which contains the 5-hydroxymethylfurfural with the mass concentration of 0.1g/Lswung dash5g/L is prepared, and reacts for 0.1hswung dash3h at 10 DEG Cswung dash60DEG C to prepare an A liquid; bovine serum albumin is dissolved in a phosphate buffer to prepare a B liquid with the bovine serum albumin concentration of 0.1g/Lswung dash10g/L; and under the stirring condition, the B liquid is added into the A liquid to react for 0.1hswung dash5h at 10 DEG Cswung dash60 DEG C, and the artificial antigen mixed liquor is obtained. The synthesis method takes the 5-hydroxymethylfurfural as hapten, and takes glutaraldehyde as a connecting arm to combine the hapten with the bovine serum albumin, so 5-hydroxymethylfurfural-bovine serum albumin is prepared, the synthesis steps are simple and effective, and the synthesis method can be used for quickly detecting the 5-hydroxymethylfurfural in food and drugs.
Description
Technical field
The present invention relates to 5 hydroxymethyl furfural complete antigen, particularly relate to a kind of synthetic method of 5 hydroxymethyl furfural-bovine serum albumin, belong to technical field of biochemical industry.
Background technology
5 hydroxymethyl furfural (having another name called 5 hydroxymethyl 2 furaldehyde, hydroxymethylfurfural, 5-HMF or 5-hydroxymethyl-furfural), English name 5-hydroxymethyl-2-furfural or 5-HMF, for needle crystal, dark yellow liquid or powder, Flos Chrysanthemi taste, there is water absorbability, soluble in water, methyl alcohol, ethanol, acetone, ethyl acetate etc.; Dissolve in ether, benzene, chloroform etc.; Be slightly soluble in tetrachloride; Being insoluble in sherwood oil, is a kind of important industrial chemicals.In its molecule, there are an aldehyde radical and a methylol, can pass through hydrogenation, oxidation deoxidation, esterification, halogenation, polymerization, hydrolysis and other chemical reaction, for the synthesis of many useful chemicals and novel high polymer material, comprise medicine, resene plastics etc.Yet, according to related documents report 5 hydroxymethyl furfural, human body voluntary muscle and internal organ are had to infringement, also be the principal element that causes nervous system disease simultaneously, its a large amount of existence meeting serious harm HUMAN HEALTH in food and medicine, the World Health Organization is controlled the content of 5 hydroxymethyl furfural in honey, and Chinese Pharmacopoeia also has very strict control to the content of 5 hydroxymethyl furfural in puerarin glucose injection liquid.Therefore, by detecting and then control the content of 5 hydroxymethyl furfural in food and medicine, have important practical significance.
At present, the method for related detection 5 hydroxymethyl furfural mainly contains both at home and abroad: colorimetry, ultraviolet method and high performance liquid chromatography (HPLC).In colorimetry, be not unique factor of glucose injection variable color due to 5 hydroxymethyl furfural, this method specificity and accuracy are poor; The concentration of ultraviolet method to sample, every country has different requirements, there is no unified standard, is difficult to determine final detected result; Use high performance liquid chromatography, although can measure very accurately the content of 5 hydroxymethyl furfural in sample, but, this method needs expensive plant and instrument and professional and technical personnel, higher to tested material requirements, sample pretreatment and operating process are complicated, and time-consuming effort again, can not meet modern measure to convenient, requirement fast and accurately.At present, the domestic report that uses immune fast detection method to detect 5 hydroxymethyl furfural content in food and medicine that there is no, in order to make up this blank, for people from now on provide material in the research aspect immunodetection, designed and take 5 hydroxymethyl furfural as the haptenic synthetic 5 hydroxymethyl furfural complete antigen that immunodetection is used that can be used for.
Summary of the invention
Shortcoming and defect for existing instrument detection technique, the preparation method who the object of this invention is to provide a kind of 5 hydroxymethyl furfural complete antigen, prepared product can be used for the immunodetection of 5 hydroxymethyl furfural content in food and medicine, for follow-up study, exploitation and application provide essential artificial antigen.
The present invention be take 5 hydroxymethyl furfural as haptens, by glutaraldehyde, is that connecting arm combines haptens and bovine serum albumin, prepares the artificial antigen of 5 hydroxymethyl furfural, i.e. 5 hydroxymethyl furfural-bovine serum albumin.
Object of the present invention is achieved through the following technical solutions.
A synthetic method for 5 hydroxymethyl furfural complete antigen, comprises the steps:
(1) preparation of A liquid: 5 hydroxymethyl furfural is dissolved in glutaraldehyde water solution, is made into the mixed solution that 5 hydroxymethyl furfural mass concentration is 0.1g/L～5g/L, react 0.1h～3h under 10 ℃～60 ℃ conditions, preparation A liquid;
(2) preparation of B liquid: bovine serum albumin is dissolved in phosphate buffered saline buffer, and being made into bovine serum albumin concentration is the B liquid of 0.1g/L～10g/L;
(3) artificial antigen is synthetic: under agitation condition, B drop being added in A liquid, is to react 0.1h～5h under 10 ℃～60 ℃ conditions in temperature, obtains artificial antigen mixed solution;
(4) dialysis: artificial antigen mixed solution is packed into and can be seen through in the dialysis tubing that molecular weight is 1kDa～100kDa, with phosphate buffered saline buffer dialysis 1 day～5 days; After dialysis finishes, dry, obtain 5 hydroxymethyl furfural complete antigen.
For further realizing the present invention, the preferred 0.1mol/L～1.0mol/L of concentration of described glutaraldehyde water solution, pH 2～7.
One or both in phosphoric acid salt preferably phosphoric acid disodium hydrogen, SODIUM PHOSPHATE, MONOBASIC and the potassium primary phosphate of described step (2) and step (4).
The concentration of the phosphate buffered saline buffer that described step (2) and step (4) are described is 0.07～0.25mol/L preferably, and pH is 7.0～7.9.
The preferred magnetic agitation of described stirring or electric stirring.
Described dry preferably freeze drying or vacuum-drying.
Compared with prior art, the present invention has the following advantages:
Productive rate is high: the connecting arm that adopts glutaraldehyde to be combined with bovine serum albumin as 5 hydroxymethyl furfural, overcome slow, the inefficient shortcoming of the direct coupling speed of 5 hydroxymethyl furfural and bovine serum albumin, greatly improved coupling speed and the efficiency of 5 hydroxymethyl furfural and bovine serum albumin, the productive rate of 5 hydroxymethyl furfural-bovine serum albumin complete antigen is improved.
Good stability: the connecting arm that adopts glutaraldehyde to be combined with bovine serum albumin as 5 hydroxymethyl furfural; farthest protected the active group in 5 hydroxymethyl furfural molecule: aldehyde radical (CHO); retain the biological activity of 5 hydroxymethyl furfural, thereby improved the stability of 5 hydroxymethyl furfural-bovine serum albumin complete antigen.
The immunodetection that can be used for 5 hydroxymethyl furfural content in food and medicine: adopt the synthetic 5 hydroxymethyl furfural-bovine serum albumin complete antigen of glutaraldehyde method, the biological activity that has farthest retained 5 hydroxymethyl furfural, so can well carry out specific binding with the corresponding antibody of 5 hydroxymethyl furfural, can be used as the use of envelope antigen in ELISA immunodetection, also can be for the capsulating material of T line in 5 hydroxymethyl furfural Rapid detection test strip, therefore can be used for the tachysynthesis of 5 hydroxymethyl furfural content in food and medicine detects, there is good application prospect.
Accompanying drawing explanation
Fig. 1 is the UV scanning figure before and after the preparation of 5 hydroxymethyl furfural complete antigen in embodiment 1.
Fig. 2 is the UV scanning figure before and after the preparation of 5 hydroxymethyl furfural complete antigen in embodiment 2.
Fig. 3 is the UV scanning figure before and after the preparation of 5 hydroxymethyl furfural complete antigen in embodiment 3.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described, but the scope of protection of present invention is not limited to the scope of embodiment statement.
Embodiment 1
The 5 hydroxymethyl furfural of 15mg is dissolved in to 5mL, and concentration is 1.0mol/L, in the glutaraldehyde water solution that pH is 2, is mixed with the mixed solution that 5 hydroxymethyl furfural mass concentration is 3.0g/L, at 30 ℃, reacts 1.5h, preparation A liquid;
The bovine serum albumin of 20mg is dissolved in to 5mL, concentration is 0.2mol/L, pH is in 7.4 phosphate buffered saline buffer (the Sodium phosphate dibasic aqueous solution configuration in 81: 19 by volume of the SODIUM PHOSPHATE, MONOBASIC of 0.2mol/L and 0.2mol/L), be mixed with the B solution that protein amount concentration is 4.0g/L, be positioned in 4 ℃ of refrigerators and preserve;
In the abundant stirring A liquid of magnetic stirring apparatus, B liquid is slowly added drop-wise in A liquid, both are fully reacted, at 10 ℃, react 5h, after having reacted, obtain artificial antigen mixed solution;
Artificial antigen mixed solution is packed into and can see through in the dialysis tubing that molecular weight is 50kDa, by concentration, be 0.2mol/L, pH is 7.4 phosphate buffered saline buffer (the Sodium phosphate dibasic aqueous solution configuration in 81: 19 by volume of the SODIUM PHOSPHATE, MONOBASIC of 0.2mol/L and 0.2mol/L) dialysis 1 day, every 12h, changes a phosphate buffered saline buffer; After dialysis finishes, artificial antigen mixed solution is carried out to lyophilize, obtain artificial antigen: 5 hydroxymethyl furfural-bovine serum albumin product.
The evaluation of 5 hydroxymethyl furfural artificial antigen: adopt ultraviolet spectrophotometer to measure its UV scanning collection of illustrative plates.UV scanning figure in the present embodiment before and after the preparation of 5 hydroxymethyl furfural complete antigen as shown in Figure 1.As can be seen from the figure there is absorption peak in 280nm left and right in bovine serum albumin, in 280nm left and right, there is absorption peak in 5 hydroxymethyl furfural, in 280nm left and right, there is a stronger absorption peak in addition, peakedness ratio bovine serum albumin is higher than the absorption peak of 5 hydroxymethyl furfural, think that this is the result of 5 hydroxymethyl furfural and the mutual superposition of bovine serum albumin success coupling postpeak, illustrates successfully synthetic 5 hydroxymethyl furfural-bovine serum albumin complete antigen.
The calculating of productive rate: productive rate is an important index evaluating a reaction combined coefficient, and this chemical reaction combined coefficient of the high explanation of productive rate is high, otherwise claims that combined coefficient is low.The quality of product (g) of the quality (g) * 100% of productive rate=actual product obtaining/in theory.After testing, it is 78.3% that the present embodiment calculates productive rate, illustrates that this law is synthesized the efficiency of 5 hydroxymethyl furfural-bovine serum albumin complete antigen higher.
Stability test: 5 hydroxymethyl furfural-bovine serum albumin complete antigen solution prepared by this law is placed in 4 ℃ of refrigerator 1-3 months, observes it and whether occurs precipitation or floss.After 3 months, observing its solution state, is still clear soln, illustrates that the synthetic 5 hydroxymethyl furfural-bovine serum albumin complete antigen of this law has good stability.
Not only productive rate is high for 5 hydroxymethyl furfural-bovine serum albumin complete antigen prepared by the present embodiment, good stability, and retained the biological activity of 5 hydroxymethyl furfural, can well carry out specific binding with the corresponding antibody of 5 hydroxymethyl furfural, can be used as the use of envelope antigen in ELISA immunodetection, also can be for the capsulating material of T line in 5 hydroxymethyl furfural Rapid detection test strip, therefore can be used for the tachysynthesis of 5 hydroxymethyl furfural content in food and medicine, detect, there is good application scenario.
The 5 hydroxymethyl furfural of 10mg is dissolved in to 10mL, and concentration is 0.1mol/L, in the glutaraldehyde water solution that pH is 5, is mixed with the mixed solution that 5 hydroxymethyl furfural mass concentration is 0.1g/L, at 10 ℃, reacts 0.1h, preparation A liquid;
The bovine serum albumin of 10mg is dissolved in to 10mL, concentration is 0.07mol/L, pH is 7.0 phosphate buffered saline buffer (configuration in 3: 2 by volume of the Sodium phosphate dibasic of 0.07mol/L and the potassium dihydrogen phosphate aqueous solution of 0.07mol/L), be mixed with the B solution that protein amount concentration is 0.1g/L, be positioned in 4 ℃ of refrigerators and preserve;
In the abundant stirring A liquid of electric mixer, B liquid is slowly added drop-wise in A liquid, both are fully reacted, at 30 ℃, react 3h, after having reacted, obtain artificial antigen mixed solution;
Artificial antigen mixed solution is packed into and can see through in the dialysis tubing that molecular weight is 10kDa, by concentration, be 0.07mol/L, pH is 7.0 phosphate buffered saline buffer (configuration in 3: 2 by volume of the Sodium phosphate dibasic of 0.07mol/L and the potassium dihydrogen phosphate aqueous solution of 0.07mol/L) dialysis 3 days, every 12h, changes a phosphate buffered saline buffer; After dialysis finishes, artificial antigen mixed solution is carried out to vacuum-drying, obtain artificial antigen: 5 hydroxymethyl furfural-bovine serum albumin product.
UV scanning figure before and after the preparation of the present embodiment 5 hydroxymethyl furfural complete antigen as shown in Figure 2.As can be seen from the figure there is absorption peak in 280nm left and right in bovine serum albumin, in 280nm left and right, there is absorption peak in 5 hydroxymethyl furfural, in 280nm left and right, find a stronger absorption peak in addition, peakedness ratio bovine serum albumin is higher than the absorption peak of 5 hydroxymethyl furfural, think that this is the result of 5 hydroxymethyl furfural and the mutual superposition of bovine serum albumin success coupling postpeak, illustrates successfully synthetic HMF-BSA complete antigen.
The productive rate that the present embodiment calculates 5 hydroxymethyl furfural-bovine serum albumin artificial antigen is 74.5%.
The 5 hydroxymethyl furfural of 25mg is dissolved in to 5mL, and concentration is 0.5mol/L, in the glutaraldehyde water solution that pH is 7, is mixed with the mixed solution that 5 hydroxymethyl furfural mass concentration is 5.0g/L, at 60 ℃, reacts 3h, preparation A liquid;
The bovine serum albumin of 100mg is dissolved in to 10mL, concentration is 0.25mol/L, pH is that the slow salt of 7.9 phosphoric acid rushes liquid (the Sodium phosphate dibasic aqueous solution of 0.25mol/L), is mixed with the B solution that protein amount concentration is 10.0g/L, is positioned in 4 ℃ of refrigerators and preserves;
In the abundant stirring A liquid of magnetic stirring apparatus, B liquid is slowly added drop-wise in A liquid, both are fully reacted, at 60 ℃, react 0.1h, after having reacted, obtain artificial antigen mixed solution;
Artificial antigen mixed solution is packed into and can see through in the dialysis tubing that molecular weight is 100kDa, by concentration, be 0.25mol/L, pH is that the slow salt of 7.9 phosphoric acid rushes liquid (the Sodium phosphate dibasic aqueous solution of 0.25mol/L) dialysis 5 days, every 12h, changes a phosphate buffered saline buffer; After dialysis finishes, artificial antigen mixed solution is carried out to vacuum-drying, obtain artificial antigen: 5 hydroxymethyl furfural-bovine serum albumin product.
UV scanning figure before and after the preparation of the present embodiment 5 hydroxymethyl furfural complete antigen as shown in Figure 3.As can be seen from the figure there is absorption peak in 280nm left and right in bovine serum albumin, in 280nm left and right, there is absorption peak in 5 hydroxymethyl furfural, in 280nm left and right, find a stronger absorption peak in addition, peakedness ratio bovine serum albumin is higher than the absorption peak of 5 hydroxymethyl furfural, think that this is the result of BSA and the mutual superposition of HMF success coupling postpeak, illustrates successfully synthetic 5 hydroxymethyl furfural-bovine serum albumin complete antigen.
The productive rate that the present embodiment calculates 5 hydroxymethyl furfural-bovine serum albumin artificial antigen is 84.5%.
5 hydroxymethyl furfural-bovine serum albumin prepared by the present invention is applied to the immuno-chromatographic test paper strip that preparation can detect 5 hydroxymethyl furfural in food and medicine, and concrete steps are as follows:
1. prepare hydroxymethylfurfural polyclonal antibody.Step 1: it is in 0.9% NaCl solution that 1mg 5 hydroxymethyl furfural-bovine serum albumin is dissolved in to 1ml massfraction, after fully emulsified with 1ml Freund's complete adjuvant again, according to the dosage of every kilogram of laboratory animal rabbit injection 0.5mg hydroxymethylfurfural-bovine serum albumin, carry out the subcutaneous multi-point injection of rabbit back; Step 2: interval is after 14 days, it is in 0.9% NaCl solution that 1mg hydroxymethylfurfural-bovine serum albumin is dissolved in to 1ml massfraction, after fully emulsified with the Freund's incomplete adjuvant of 1ml again, dosage according to every kilogram of laboratory animal rabbit injection 0.5mg hydroxymethylfurfural-bovine serum albumin carries out the subcutaneous multi-point injection of rabbit back, then every 14 days repeating steps 21 times, repeat altogether 3 times.Last immunity, after ten days, is carried out taking blood from jugular vein to rabbit, separation of serum, and with saturated ammonium sulphate method precipitating antibody, and with affinity chromatography purifies and separates hydroxymethylfurfural antibody.By indirect enzyme-linked immunosorbent assay (ELISA) method, detecting many anti-tiring is 1: 128000.
2. fluorescent latex mark hydroxymethylfurfural antibody and rabbit igg.Fluorescent latex mark hydroxymethylfurfural antibody and rabbit immunoglobulin G (rabbit igg).(color is for yellow to get 0.1ml mass concentration and be 10% fluorescent latex microballoon, particle diameter is 0.5 μ m, excitation wavelength is 600nm), with 0.9ml, the MES damping fluid of 0.1mol/LpH 6 is adjusted into 1% by the concentration of fluorescent latex microballoon, then order adds 5mgEDC and 5mg Sulfo-N HS activation, mild stirring 30min under room temperature, after activation finishes, solution is centrifugal 30min under 1000rpm rotating speed, precipitation is dissolved with the PBS damping fluid of 0.1mol/L pH 6, through ultrasonic resuspended after, adding respectively 40 μ l mass concentrations is 10mg/ml rabbit igg (ZI103-1 rabbit igg, the true Bioisystech Co., Ltd in Shanghai) and the hydroxymethylfurfural polyclonal antibody 1. prepared of step, after vibrator evenly mixes, mild stirring reaction 3h under room temperature, after reaction finishes, solution is centrifugal 30min under 1000rpm rotating speed, precipitation by PBS damping fluid (the 0.1mol/L pH 7.0) repeated washing three times that is dissolved with 0.05%Tween 20 to remove the albumen on unmarked, centrifugal 30min under 1000rpm rotating speed, precipitation 0.1mol/L, pH 7.4 is dissolved with 1% bovine serum albumin and 0.05%Na simultaneously
3the PBS damping fluid of N dissolves again, through ultrasonic resuspended after, be positioned in 4 ℃ of refrigerators stand-byly, through the hydroxymethylfurfural antibody of fluorescent latex mark and the final concentration of rabbit igg, be now 0.3mg/ml.
3. prepare fluorescent latex pad.With 0.1mol/L, pH 7.4 is dissolved with 1% bovine serum albumin and 0.05%Na simultaneously
3fluorescently-labeled hydroxymethylfurfural polyclonal antibody and rabbit igg that 2. the PBS damping fluid of N is prepared step dilute 100 times simultaneously, the ratio that is in mass ratio 1: 500 is evenly mixed, its mixed solution is evenly sprayed on trevira film (the SB06 Shanghai biological company limited of gold mark), trevira film is placed in to dry 36h under room temperature, standby.
4. prepare coated film.With 0.1mol/L, pH is that 7.4 PBS damping fluid is adjusted into 2mg/ml by hydroxymethylfurfural-protein amount concentration, the mass concentration of goat anti-rabbit igg (the true Bioisystech Co., Ltd in Shanghai) is adjusted into 0.25mg/ml, hydroxymethylfurfural complete antigen and goat anti-rabbit igg are evenly sprayed at respectively to the different zones on nitrocellulose filter, form banded detection zone and Quality Control district, wherein detection zone and Quality Control interval are every 5mm, coated film is positioned over to dry 36h in 50 ℃ of baking ovens, standby.
5. prepare immuno-chromatographic test paper strip.Take vinyon plate as support base, by order from left to right, in turn sample pad is overlapped in to distance fluorescent latex pad leftmost edge 2mm place on fluorescent latex pad, fluorescent latex pad is overlapped in coated film, and (detection zone is on a left side, Quality Control district is on the right side) upper distance coated film leftmost edge 2mm place, thieving paper is overlapped in 2mm place, edge, distance coated film the right on coated film, and it is wide to cut into 0.4cm, the test strip that 7.7cm is long.
Content by the immuno-chromatographic test paper strip of preparation for detection of 5 hydroxymethyl furfural in food and medicine, result shows, the measurement result of this immuno-chromatographic test paper strip and the measurement result of liquid phase chromatography approach, time is only 5min, illustrates that the synthetic 5 hydroxymethyl furfural complete antigen of this law can be used for the tachysynthesis detection of 5 hydroxymethyl furfural in food and medicine.
Claims (4)
1. a synthetic method for 5 hydroxymethyl furfural complete antigen, is characterized in that comprising the steps:
(1) preparation of A liquid: by 5 ?hydroxymethylfurfural be dissolved in glutaraldehyde water solution, be made into 5 ?the hydroxymethylfurfural mass concentration mixed solution that is 0.1g/L～5g/L, under 10 ℃～60 ℃ conditions, react 0.1h～3h, preparation A liquid; The concentration of described glutaraldehyde water solution is 0.1mol/L～1.0mol/L, and pH is 2～7;
(2) preparation of B liquid: bovine serum albumin is dissolved in phosphate buffered saline buffer, and being made into bovine serum albumin concentration is the B liquid of 0.1g/L～10g/L;
(3) artificial antigen is synthetic: under agitation condition, B drop being added in A liquid, is to react 0.1h～5h under 10 ℃～60 ℃ conditions in temperature, obtains artificial antigen mixed solution;
(4) dialysis: artificial antigen mixed solution is packed into and can be seen through in the dialysis tubing that molecular weight is 1kDa～100kDa, with phosphate buffered saline buffer dialysis 1 day～5 days; After dialysis finishes, dry, obtain 5 ?hydroxymethylfurfural complete antigen;
The concentration of the phosphate buffered saline buffer that step (2) and step (4) are described is 0.07～0.25mol/L, and pH is 7.0～7.9.
According to claim 1 a kind of 5 ?the synthetic method of hydroxymethylfurfural complete antigen, it is characterized in that: the phosphoric acid salt of described step (2) and step (4) is Sodium phosphate dibasic and SODIUM PHOSPHATE, MONOBASIC.
According to claim 1 a kind of 5 ?the synthetic method of hydroxymethylfurfural complete antigen, it is characterized in that: described stirring is magnetic agitation or electric stirring.
According to claim 1 a kind of 5 ?the synthetic method of hydroxymethylfurfural complete antigen, it is characterized in that: described being dried as lyophilize or vacuum-drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110228269.1A CN102382189B (en) | 2011-08-10 | 2011-08-10 | Synthesis method of 5-hydroxymethylfurfural complete antigen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110228269.1A CN102382189B (en) | 2011-08-10 | 2011-08-10 | Synthesis method of 5-hydroxymethylfurfural complete antigen |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102382189A CN102382189A (en) | 2012-03-21 |
| CN102382189B true CN102382189B (en) | 2014-04-02 |
Family
ID=45822073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110228269.1A Active CN102382189B (en) | 2011-08-10 | 2011-08-10 | Synthesis method of 5-hydroxymethylfurfural complete antigen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102382189B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102675454B (en) * | 2012-05-09 | 2014-04-02 | 华南理工大学 | Synthetic method of 4-methylimidazole complete antigen |
| CN109307761B (en) * | 2018-10-09 | 2021-06-15 | 华南农业大学 | Indirect competitive ELISA method for detecting furaldehyde |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080268477A1 (en) * | 2005-04-05 | 2008-10-30 | Jms Co., Ltd. | Antibody Reactive Specifically to Age Derived from 3,4-Dge |
-
2011
- 2011-08-10 CN CN201110228269.1A patent/CN102382189B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN102382189A (en) | 2012-03-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102384973B (en) | Immunochromatography test strip for quickly and quantitatively detecting hydroxymethylfurfural and preparation method thereof | |
| CN103575893A (en) | Method for rapidly detecting shellfish toxin | |
| CN102590514B (en) | Method for detecting illegal cooking oil, test paper and application of test paper | |
| CN1314707C (en) | 2, 4, 6-trichlorophen artificial antigen, and its preparing method and use | |
| CN102323428A (en) | Riemerella anatipestifer antibody indirect ELISA method detection kit and application thereof | |
| CN101245032A (en) | Leuco malachite green hapten, produced antibody and application of the antibody | |
| CN103613563B (en) | Preparation method of benzothiostrobin hapten, artificial antigen and specific antibody and application thereof | |
| CN103360488A (en) | Preparation method for artificial antigen of theophylline | |
| CN104327183A (en) | Preparation method and application of paclobutrazol artificial antigen and polyclonal antibody | |
| CN106866568A (en) | Furaxone metabolite AOZ derivatizations haptens, the preparation method and applications of artificial antigen | |
| CN101830980B (en) | Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method | |
| CN102382189B (en) | Synthesis method of 5-hydroxymethylfurfural complete antigen | |
| CN100478357C (en) | Ofloxacin couple and its preparing method and use | |
| CN102898410B (en) | Method for preparing anti-coagulation raticide bromadiolone half antigen and complete antigen | |
| CN101781365A (en) | Artificial antigen of tetraodotoxin and corresponding specific antibody and preparation method and application thereof | |
| CN101173007B (en) | 1-amido-glycolyurea immunogen, producing method and application of the same | |
| CN102875512B (en) | Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen | |
| CN105399639A (en) | Tyramine artificial antigen and antibody, and preparation methods and application thereof | |
| CN103288741A (en) | Histamine hapten, artificial antigen, antibody, as well as preparation methods and applications thereof | |
| CN104977401A (en) | Purifying preparation method of allergen positive serum | |
| CN106831498A (en) | Furacilin metabolite SEM derivatizations haptens, the preparation method and applications of artificial antigen | |
| CN104744294B (en) | (α ethyl hexyls) ester hapten, artificial antigen and its preparation method for antibody of phthalic acid two and application | |
| CN1904613B (en) | Preparation method of artificial antigen used for immune analysis of sulfanilamide multi kind residue | |
| CN105085665A (en) | Vitamin B2 conjugate and preparation method and application thereof | |
| CN103364546B (en) | A kind of kit and method detecting Furaxone metabolite |
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 |