CN102898515A - Polybrominated biphenyls homologue immunogen and preparation method - Google Patents
Polybrominated biphenyls homologue immunogen and preparation method Download PDFInfo
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Abstract
The invention relates to a polybrominated biphenyls homologue immunogen and a preparation method, which comprises the following steps: dissolving succinic anhydride and 4,4'-dibromobiphenyl in dichloromethane, stirring in ice bath, feeding anhydrous aluminum trichloride under the condition of ice water-bath, stirring in a seal mode and adding dilute hydrochloric acid for completing a reaction, processing to obtain PBB28 semiantigen 4-(4,4'-dibromobiphenyl)-4oxygen-butyric acid, then mixing N-hydroxy succinimide and N,N'-dicyclohexylcarbodiimide, adding bovine serum protein for stirring and reacting and then transferring to a semi-permeable membrane, dialyzing in a phosphatic buffer solution, centrifuging to obtain a supernatant to obtain a PBB28 artificial immunogen PBB-BSA conjugate. The method of the invention has the advantages of simple preparation method, low cost and easy industrial production; the polybrominated biphenyls artificial immunogen PBB28-BSA can prepare a specific antibody having an immunization reaction characteristic on a polybrominated biphenyls monomer through immunization animals, and the polybrominated biphenyls homologue immunogen can be used for on-site detection of the polybrominated biphenyls monomer with trace amount in water, soil and atmosphere.
Description
Technical field
The present invention relates to a kind of immunogen and preparation method thereof, especially relate to a kind of Polybrominated biphenyl homologue immunogen and preparation method thereof.
Background technology
Bromide fire retardant is one of organic fire-retardant of present in the world output maximum, wherein mainly be Poly Brominated Diphenyl Ethers (PBDEs) and Polybrominated biphenyl (PBBs) class material, Chang Zuowei is fire-retardant to be added in the Polymer Synthesizing materials such as agent adding resin, polystyrene and polyurethane foam, be widely used in the fields such as plastics, textiles, circuit card and material of construction, and along with the use of these products, slowly spill in the environment.
Polybrominated biphenyl and polychlorobiphenyl chemical property are similar, are a kind of highly stable and easily form the chemical of biological accumulation in environment, also grow a lot in these several years about the understanding of the bio-toxicity of Polybrominated biphenyl.By PBBs and PBDEs cumulative effect and the environment, the food contamination problem that cause have caused that the developed countries such as European Union, the U.S. show great attention to, the related objectionable impurities of restriction instruction (ROHS) instruction of objectionable impurities comprises fire retardant (Flame Retardants)-Polybrominated biphenyl (PBBs) and Poly Brominated Diphenyl Ethers (PBDEs) in European Union's electronic and electrical equipment.At present environmental protection organization mainly concentrates on the toxicity aspect of the environmental area such as water body, soil, atmosphere and organism about the research of PBDE and PBBs, and all there are larger difference in determination process, scavenging process and analytic process.Therefore, conduct a research and set up effectively, reliably detection method and the standard of brominated flame-retardant, and ensured by relevant rules, become a very important and urgent job.
Immunoassay is based on antigen and antibody specific identification and association reaction is basic analytical procedure.Immunoassay has high specificity, highly sensitive (detectability can reach 10
-6～10
-9G), convenient and swift, the analysis capacity is large, analysis cost is low, the advantage such as safe and reliable.Generally do not need expensive instrument, can greatly simplify even save pretreatment process, less demanding to user of service's technical skill, easily popularize and promote.
For the foundation of the immunologic surveillance method of Polybrominated biphenyl, domesticly also do not see up to now relevant report.This may be because Polybrominated biphenyl homologue of a great variety, and, the physics of PBBs homologue, chemistry, biology and toxicological different in kind.Therefore relevant forward position research requires the analysis of PBBs must be by the level rise of measuring total amount on the level of mensuration congener and isomer, so the qualitative and quantitative analysis of single PBBs homologue is very necessary.
Therefore, the preparation of Polybrominated biphenyl homologue and derivative thereof is to its toxicology character, Transport And Transformation, and the research such as ecologic effect is significant.Polybrominated biphenyl haptens and immunogenic preparation are to adopt immunization method to measure the basis of Polybrominated biphenyl, for the monitoring method of setting up the Polybrominated biphenyl monomer, have important using value and theoretical significance.
The Polybrominated biphenyl monomer of selling on the Vehicles Collected from Market is few, more do not see about can and macro-molecular protein form immunogenic Polybrominated biphenyl hapten derivant molecule, therefore, the research about the immunologic detection method of polybrominated biphenyl environmental pollutant yet there are no report so far.For the foundation of Polybrominated biphenyl immunologic surveillance method, the haptens that high purity is suitable is that preparation has the basic of highly sensitive and specific immunogens and high-titer antibody.
Summary of the invention
Purpose of the present invention is exactly to provide a kind of step simple for the defective that overcomes above-mentioned prior art existence, and speed is fast, the much higher bromo biphenyl homologue of productive rate immunogen and preparation method thereof.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of Polybrominated biphenyl homologue immunogen, this immunogenic structural formula is as follows:
The immunogenic preparation method of a kind of Polybrominated biphenyl homologue may further comprise the steps:
(1) PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid is synthetic:
Syntheti c route is as follows:
With Succinic anhydried, 4,4 '-'-dibromobiphenyl is dissolved in the non-protonic solvent, ice bath stirs, aluminum trichloride (anhydrous) feeds intake under the ice-water bath condition, and gradation adds in 1-2h, and aluminum trichloride (anhydrous) dropwises the water-bath of recession deicing, under normal temperature condition, 10-14h is stirred in the water proof sealing, and thin-layer method monitoring reaction raw material point adds the dilute hydrochloric acid termination reaction after disappearing.Organic layer is removed organic solvent with dilute hydrochloric acid washing and rotary evaporation, gets yellow solid powder crude product, through recrystallization, drying, obtains PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid;
(2) preparation of PBB28 artificial immunogen PBB28-BSA:
Syntheti c route is as follows:
Under 0-5 ℃ condition, to be dissolved with N-hydroxy-succinamide and N, the organic solution of N '-dicyclohexyl carbimide dropwise adds and is dissolved with in the haptenic organic solution of PBB28, stirring reaction 6-12h, 4 ℃ are spent the night, it is 6.8-7.4 that the centrifugal upper strata Acibenzolar of telling is joined pH, be dissolved with in the phosphate buffer soln of bovine serum albumin (BSA), react 4-6h in 0-4 ℃ the ice bath, the centrifugal supernatant liquor that obtains, supernatant liquor is transferred in the semi-permeable membranes, be the phosphate buffer soln of 6.8-7.4 or 0.9% normal saline dialysis 5 days at pH, namely get PBB28 artificial immunogen (PBB-BSA conjugate), purify through post, add ℃ lower or freeze-drying preservation of equivalent glycerine-20.
Non-proton organic solvent described in the step (1) is methylene dichloride or oil of mirbane.
Described in the step (1) 4,4 '-'-dibromobiphenyl, Succinic anhydried and anhydrous AlCl
3Mol ratio be 1: 1-2: 1-2.5.
It is that the mixing solutions that 1: 1 methyl alcohol and ether form carries out recrystallization that step (1) adopts volume ratio.
The solvent that adopts in the described organic solution of step (2) is N ' dinethylformamide or dimethyl sulfoxide (DMSO).
The described PBB28 haptens of step (2), N-hydroxy-succinamide, N, the mol ratio of N '-dicyclohexyl carbimide and bovine serum albumin is 1: 1-4: 1-4: 0.005-0.05.
The sign of holoantigen
The holoantigen of preparation is suitably diluted, make its absorbancy between 0.1-2, take PBS as blank, measure respectively '-dibromobiphenyl (PBB28) with ultraviolet spectrophotometer at 200～500nm, BAS, with the light absorption value of PBB28-BSA, draw uv absorption spectra, and calculate coupling ratio according to following formula:
Compared with prior art, the present invention has the following advantages:
(1) adopts single step reaction, the carboxyl functional group that contains 4 carbon atoms is introduced in position according to the bromine atoms on the raw material Polybrominated biphenyl at phenyl ring, make corresponding Polybrominated biphenyl homologue semiantigen, then haptens is passed through active ester method, be connected respectively to (BSA or OVA) on the carrier protein, carboxyl on the haptens generates an intermediate with N-hydroxy-succinamide (NHS) reaction first, and then with protein on amino coupled, form the haptens protein conjugate, i.e. the artificial immunogen of Polybrominated biphenyl (perhaps coating antigen)
(2) product after degrading under field conditions (factors) for Polybrominated biphenyl in the environment mostly is this situation of low bromo biphenyl that contains 3-4 bromine; at first adopt the Friedel-Crafts acylation reaction; choose 4; the carboxyl that contains four carbon has been introduced at the ortho position of bromine atoms on 4 '-'-dibromobiphenyl (PBB28) phenyl ring; prepare corresponding haptens; then adopt active ester method macro-molecular protein and hapten conjugation, prepare corresponding antigen.
(3) the reagent price of using is low, and the reaction times is short, and the method aftertreatment is simple to operation, is suitable for the antigen preparation of other Polybrominated biphenyl homologues.
Description of drawings
Fig. 1 is '-dibromobiphenyl haptens infrared spectra;
Fig. 2 is '-dibromobiphenyl antigen uv absorption spectra;
Fig. 3 is the real-time fluorescence immuno-PCR amplification curve of Direct Determination Polybrominated biphenyl;
Fig. 4 is for measuring the standard working curve of PBB28.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
(1) 4, the preparation of 4 '-'-dibromobiphenyl haptens
Take by weighing 4,4 '-'-dibromobiphenyl 5mmol is dissolved in 20mL CH is housed
2Cl
2Flask in, add the succinyl oxide of 5mmol, stirring and dissolving under the condition of ice bath, adds the anhydrous AlCl of 12.5mmol
3Gradation adds in 1.5h.After 10h was stirred in sealing, stopped reaction in the hydrochloric acid of reaction solution impouring 100ml 2M, was then used the CH of 50ml * 3
2Cl
2Extraction with the salt acid elution extraction liquid of 100ml 2M, is used anhydrous magnesium sulfate drying, filters, and removes solvent, gets crude product.With 1: 1 methyl alcohol and diethyl ether solution recrystallization, obtain pink powdered granule, survey fusing point: 162-164 ℃.Product verifies with ultimate analysis through infrared spectra and identifies, the result is: IR, σ/cm
-1: 3395 (OH), 2956,2927 (CH
2), 1709 (C=O), 1589,1375 (VS, CO2
-), 1134 (C-Br), 883,822 (C-H).Results of elemental analyses, calculated value (measured value), %:C 46.24 (46.60), H 2.716 (2.912) .N 0 (＜0.030);
Find out from the infared spectrum analysis of Fig. 1, at 3395cm
-1The hydroxyl peak of carboxyl has appearred in the place, at 2956cm
-1The place has the C-H shock absorbing peak of methyl 1589cm to occur
-1(s, CO2
-) and 1375 (VS, CO2
-) cm
-1Two carbonyl vibration performance peaks that the peak is carboxyl, thus illustrate that the carbochain with carboxyl is connected on the phenyl ring.Results of elemental analyses has proved that further the structure of product is consistent with designed route.
(2) preparation of immunizing antigen
The Polybrominated biphenyl haptens of the present invention's preparation is mainly used in the immunodetection of Polybrominated biphenyl in the environment.One of its main application can be used for directly and the protein macromolecule coupling exactly, prepares the immunogen of immune animal, and then prepares corresponding mono-clonal or polyclonal antibody.Set up on this basis the method for immunity of Polybrominated biphenyl.The below is with regard to 4, and immunogenic the making of 4 '-'-dibromobiphenyl is used as with applicating example:
Take by weighing 0.4mmol haptens 4-(4,4 '-'-dibromobiphenyl base)-4-oxygen-butyric acid, place the fine taper bottle of clean dried, add 400 μ L DMF and make it dissolving.Take by weighing 64.8mg DCC and 38.8mg NHS and be dissolved in 300 μ LDMF, under magnetic agitation, dropwise be added in the fine taper bottle, stirring at room reaction 8h, 4 ℃ are spent the night.To react after product and place low-temperature and high-speed whizzer 9000r/min, isolate supernatant liquor through 15min.Take by weighing 160mgBSA and be dissolved in 10mL 0.02mol/L pH 7.4 phosphate buffer solns, dropwise add the above-mentioned clear liquid of 600 μ L, under condition of ice bath, react 4h.Reaction is finished, with the solution dialysis tubing of packing into, and tap water flushing 48h,, 0.9% normal saline dialysis 3 days, the centrifugal supernatant liquor of telling namely gets the PBB-BSA conjugate.Conjugate can be seen from the uv absorption spectra 2 of conjugate PBB-BSA, from PBB after ultraviolet-visible pectrophotometer scanning is identified
2(B line), BSA (C line), and conjugate PBB
2The uv absorption spectra of-BSA (D line) can be seen, 4-(4,4 '-the '-dibromobiphenyl base)-4 oxygen-butyric acid has maximum absorption band at 283 wavelength places, BSA has maximum absorption band at wavelength 280nm place, reach appears in the highest absorption peak of PBB-BSA, meets PBB and BSA absorption peak Superposition Characteristics.Calculate the combination of PBB and BSA among the PBB-BSA than being respectively 19: 1.
These immunogens both can be used for immune animal, and the immune response that produces by animal obtains can being used for doing mono-clonal or the polyclonal antibody of immunoassay, can be used as again the Immune competition object and used.
Embodiment 2
The animal immune experimental verification
Select 2 adult healthy male New Zealand rabbits, the holoantigen that synthesizes is through with after Freund's complete adjuvant fully mixes, White Rabbit is carried out neck and back point-like injecting immune, through behind 7 booster immunizations, test tube method and agar double diffusion test are investigated its antiserum titre and have been reached 1: 320 respectively (1: 32) and 1: 640 (1: 64), agar double diffusion test shows that cross reaction is all not obvious, illustrates that its specificity is better.
Embodiment 3
The real-time Immunal PCR of direct competitive is measured PBBs
Get the polypropylene PCR pipe of clean 0.2mL specification, 0.8% concentration glutaraldehyde solution, 50 μ L, process 5-6h by 37 ℃.Ultrapure water 3min * 3 time are cleaned.With coated damping fluid (CBS) dilution PBB-OVA solution, be added in the PCR tubule of processing through glutaraldehyde (20 μ L/ hole), 4 ℃ are spent the night, and wash with washings (PBST) and manage 3min * 3 time.Every pipe adds the PBS solution that 200 μ L contain 3%OVA, and 37 ℃ of incubation 1h wash pipe 3min * 3 time with PBST.Add 25 μ LPBB28 small molecules and the anti-PBB28 polyclonal antibody of 25 μ L biotinylations in same PCR tubule, 37 ℃ of incubation 1h wash pipe 3min * 3 time with PBST.Every pipe adds the avidin solution of 50 μ L, 6.5 μ g/mL, 37 ℃ of incubation 0.5h.With PBST washing three times, each 3min.Every pipe adds the biotinylated DNA of 50 μ L, and behind 37 ℃ of incubation 1h, first with PBST washing 5 times, then with ultrapure water washing five times, each 3min. adds amplimer and fluorescent substance at last, increases in the Fluorescence PCR instrument.The Ct value of measuring and the amount of determined antigen are inversely proportional to.Make typical curve according to the antigen that adds concentration known and corresponding Ct value, thereby can obtain the concentration of PBBs in the corresponding testing sample.The amplification curve of its mensuration and the typical curve of foundation are as shown in Figure 3 and Figure 4.
Embodiment 4
The immunogenic preparation method of a kind of Polybrominated biphenyl homologue may further comprise the steps:
(1) PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid is synthetic:
With Succinic anhydried, 4,4 '-'-dibromobiphenyl is dissolved in the methylene dichloride, and ice bath stirs, and aluminum trichloride (anhydrous) feeds intake under the ice-water bath condition, and gradation adds in 1h, and 4,4 '-'-dibromobiphenyl, Succinic anhydried and anhydrous AlCl
3Mol ratio be 1: 1: 1, aluminum trichloride (anhydrous) dropwises the water-bath of recession deicing, under normal temperature condition, after adding the dilute hydrochloric acid termination reaction behind the water proof sealing stirring 10h, organic layer is removed organic solvent with dilute hydrochloric acid washing and rotary evaporation, gets the yellow solid powder, adopting volume ratio is that the mixing solutions that 1: 1 methyl alcohol and ether form carries out recrystallization, drying obtains PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid;
(2) preparation of PBB28 artificial immunogen PBB28-BSA:
Under 0 ℃ condition, to be dissolved with N-hydroxy-succinamide and N, N ' the dinethylformamide of N '-dicyclohexyl carbimide dropwise adds and is dissolved with in the haptenic N ' dinethylformamide of PBB28, stirring reaction 6h, 4 ℃ are spent the night, it is 6.8 that the centrifugal upper strata Acibenzolar of telling is joined pH, be dissolved with in the phosphate buffer soln of bovine serum albumin (BSA), the PBB28 haptens, N-hydroxy-succinamide, N, the mol ratio of N '-dicyclohexyl carbimide and bovine serum albumin is 1: 1: 1: 0.005, reaction 2h obtains supernatant liquor in 0 ℃ the ice bath, supernatant liquor is transferred in the semi-permeable membranes, and be 6.8 phosphate buffer soln dialysis 5 days at pH, the centrifugal supernatant liquor of telling, namely get PBB28 artificial immunogen PBB-BSA conjugate, purify through post, add ℃ lower or freeze-drying preservation of equivalent glycerine-20.
The immunogenic preparation method of a kind of Polybrominated biphenyl homologue, the method may further comprise the steps:
(1) PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid is synthetic:
With Succinic anhydried, 4,4 '-'-dibromobiphenyl is dissolved in the oil of mirbane, and ice bath stirs, and aluminum trichloride (anhydrous) feeds intake under the ice-water bath condition, and gradation adds in 2h, and 4,4 '-'-dibromobiphenyl, Succinic anhydried and anhydrous AlCl
3Mol ratio be 1: 2: 2.5, aluminum trichloride (anhydrous) dropwises the water-bath of recession deicing, under normal temperature condition, after adding the dilute hydrochloric acid termination reaction behind the water proof sealing stirring 14h, organic layer is removed organic solvent with dilute hydrochloric acid washing and rotary evaporation, gets the yellow solid powder, adopting volume ratio is that the mixing solutions that 1: 1 methyl alcohol and ether form carries out recrystallization, drying obtains PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid;
(2) preparation of PBB28 artificial immunogen PBB28-BSA:
Under 5 ℃ condition, to be dissolved with N-hydroxy-succinamide and N, the dimethyl sulphoxide solution of N '-dicyclohexyl carbimide dropwise adds and is dissolved with in the haptenic dimethyl sulphoxide solution of PBB28, stirring reaction 12h, 4 ℃ are spent the night, it is 7.4 that the centrifugal upper strata Acibenzolar of telling is joined pH, be dissolved with in the phosphate buffer soln of bovine serum albumin (BSA), the PBB28 haptens, N-hydroxy-succinamide, N, the mol ratio of N '-dicyclohexyl carbimide and bovine serum albumin is 1: 4: 4: 0.05, reaction 6h obtains supernatant liquor in 4 ℃ ice bath, supernatant liquor is transferred in the semi-permeable membranes, 0.9% normal saline dialysis 5 days, the centrifugal supernatant liquor of telling, namely get PBB28 artificial immunogen PBB-BSA conjugate, purify through post, add ℃ lower or freeze-drying preservation of equivalent glycerine-20.
Claims (7)
1. a Polybrominated biphenyl homologue immunogen is characterized in that, this immunogenic structural formula is as follows:
2. the immunogenic preparation method of Polybrominated biphenyl homologue as claimed in claim 1 is characterized in that, the method may further comprise the steps:
(1) PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid is synthetic:
With Succinic anhydried, 4,4 '-'-dibromobiphenyl is dissolved in the non-protonic solvent, ice bath stirs, aluminum trichloride (anhydrous) feeds intake under the ice-water bath condition, gradation adds in 1-2h, aluminum trichloride (anhydrous) adds the water-bath of recession deicing, under normal temperature condition, after water proof sealing was stirred and added the dilute hydrochloric acid termination reaction behind the 10-14h, organic layer was removed impurity with the dilute hydrochloric acid washing, rotary evaporation is removed organic solvent, get yellow solid powder crude product, recrystallization, drying, obtain PBB28 haptens 4-(4,4 '-'-dibromobiphenyl base)-4 oxygen-butyric acid;
(2) preparation of PBB28 artificial immunogen PBB28-BSA:
Under 0-5 ℃ condition, to be dissolved with N-hydroxy-succinamide and N, the organic solution of N '-dicyclohexyl carbimide dropwise adds and is dissolved with in the haptenic organic solution of PBB28, stirring reaction 6-12h, 4 ℃ are spent the night, it is 6.8-7.4 that the centrifugal upper strata Acibenzolar of telling is joined pH, be dissolved with in the phosphate buffer soln of bovine serum albumin (BSA), stirring reaction 4-6h obtains supernatant liquor in 0-4 ℃ the ice bath, supernatant liquor is transferred in the semi-permeable membranes, be the phosphate buffer soln of 6.8-7.4 or 0.9% normal saline dialysis 5 days at pH, the centrifugal supernatant liquor of telling, namely get PBB28 artificial immunogen PBB-BSA conjugate, purify through post, add equivalent glycerine-20 ℃ lower the preservation or the freeze-drying preservation.
3. the immunogenic preparation method of Polybrominated biphenyl homologue according to claim 2 is characterized in that, the non-proton organic solvent described in the step (1) is methylene dichloride or oil of mirbane.
4. the immunogenic preparation method of Polybrominated biphenyl homologue according to claim 2 is characterized in that, 4 described in the step (1), 4 '-'-dibromobiphenyl, Succinic anhydried and anhydrous AlCl
3Mol ratio be 1: 1-2: 1-2.5.
5. the immunogenic preparation method of Polybrominated biphenyl homologue according to claim 2 is characterized in that, it is that the mixing solutions that 1: 1 methyl alcohol and ether form carries out recrystallization that step (1) adopts volume ratio.
6. the immunogenic preparation method of Polybrominated biphenyl homologue according to claim 2 is characterized in that, the solvent that adopts in the described organic solution of step (2) is N ' dinethylformamide or dimethyl sulfoxide (DMSO).
7. the immunogenic preparation method of Polybrominated biphenyl homologue according to claim 2, it is characterized in that, the described PBB28 haptens of step (2), N-hydroxy-succinamide, N, the mol ratio of N '-dicyclohexyl carbimide and bovine serum albumin is 1: 1-4: 1-4: 0.005-0.05.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103288965A (en) * | 2013-06-20 | 2013-09-11 | 重庆理工大学 | Polychlorobiphenyl monoclonal antibody preparation method |
| CN103698506A (en) * | 2013-12-31 | 2014-04-02 | 杭州爱贝亚检测技术有限公司 | Method for detecting content of pollutants in sample by utilizing direct immuno-PCR (polymerase chain reaction) assay |
| CN108752463A (en) * | 2018-05-29 | 2018-11-06 | 上海交通大学 | A kind of preparation method and its usage of 2,4,4 '-tribromo Biphenyl Ether immunogene |
| CN111320698A (en) * | 2020-03-26 | 2020-06-23 | 江西省农业科学院农产品质量安全与标准研究所 | Preparation method and application of CdTe labeled antibody and method for detecting polybrominated biphenyls |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101157611A (en) * | 2007-11-16 | 2008-04-09 | 东华大学 | Polychlorinated biphenyl (PCBs) homologue semiantigen and preparation method thereof |
| CN101781050A (en) * | 2010-03-09 | 2010-07-21 | 江汉大学 | Method for treating low-concentration chromate waste water |
-
2012
- 2012-08-08 CN CN2012102808983A patent/CN102898515A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101157611A (en) * | 2007-11-16 | 2008-04-09 | 东华大学 | Polychlorinated biphenyl (PCBs) homologue semiantigen and preparation method thereof |
| CN101781050A (en) * | 2010-03-09 | 2010-07-21 | 江汉大学 | Method for treating low-concentration chromate waste water |
Non-Patent Citations (4)
| Title |
|---|
| 《化学试剂》 20081231 陈寒玉等 多溴联苯的免疫检测研究I-半抗原的制备和表征 第203-205页 1-7 第30卷, 第3期 * |
| 曹婉真, 夏又新: "《电解质》", 30 June 1991, article "1.6.6 溶剂的分类", pages: 35 * |
| 陈寒玉等: "多溴联苯免疫PCR人工抗原的合成及表征", 《环境化学》, vol. 27, no. 5, 30 September 2008 (2008-09-30), pages 621 - 623 * |
| 陈寒玉等: "多溴联苯的免疫检测研究I—半抗原的制备和表征", 《化学试剂》, vol. 30, no. 3, 31 December 2008 (2008-12-31), pages 203 - 205 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103288965A (en) * | 2013-06-20 | 2013-09-11 | 重庆理工大学 | Polychlorobiphenyl monoclonal antibody preparation method |
| CN103698506A (en) * | 2013-12-31 | 2014-04-02 | 杭州爱贝亚检测技术有限公司 | Method for detecting content of pollutants in sample by utilizing direct immuno-PCR (polymerase chain reaction) assay |
| CN103698506B (en) * | 2013-12-31 | 2015-07-29 | 杭州爱贝亚检测技术有限公司 | Direct immunization PCR is utilized to detect the method for pollutant load in sample |
| CN108752463A (en) * | 2018-05-29 | 2018-11-06 | 上海交通大学 | A kind of preparation method and its usage of 2,4,4 '-tribromo Biphenyl Ether immunogene |
| CN111320698A (en) * | 2020-03-26 | 2020-06-23 | 江西省农业科学院农产品质量安全与标准研究所 | Preparation method and application of CdTe labeled antibody and method for detecting polybrominated biphenyls |
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