Polybrominated biphenyls homologue immunogen and preparation method

A polybrominated biphenyl and immunogen technology, which is applied in the field of polybrominated biphenyl homologue immunogen and its preparation, can solve the problems that the immunological detection method of polybrominated biphenyl environmental pollutants has not been reported, and the polybrominated biphenyl hapten derivative molecules have not been seen, etc., to achieve post-processing Simple and easy operation, short reaction time and low price

Inactive Publication Date: 2013-01-30
SHANGHAI JIAO TONG UNIV
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are not many PBB monomers currently on the market, and there are no PBB hapten derivative molecules that can form immunogens with macromolecular proteins. Therefore, research on immunological detection methods for PBB environmental pollutants has not yet been seen. to report

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Polybrominated biphenyls homologue immunogen and preparation method
  • Polybrominated biphenyls homologue immunogen and preparation method
  • Polybrominated biphenyls homologue immunogen and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Preparation of 4,4'-dibromobiphenyl hapten

[0040] Weigh 5mmol of 4,4'-dibromobiphenyl, dissolve in 20mL CH 2 Cl 2 In the flask, add 5mmol of succinic anhydride, stir to dissolve, under the condition of ice bath, add 12.5mmol of anhydrous AlCl 3 . Added in portions within 1.5h. After sealing and stirring for 10 hours, stop the reaction, pour the reaction solution into 100ml 2M hydrochloric acid, and then use 50ml×3 CH 2 Cl 2 For extraction, wash the extract with 100ml of 2M hydrochloric acid, dry over anhydrous magnesium sulfate, filter, and remove the solvent to obtain a crude product. Recrystallize with 1:1 methanol and ether solution to obtain pink powdery particles. Melting point: 162-164°C. The product is verified by infrared spectroscopy and identified by elemental analysis. 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). Elemental analysis results, calculated value (measu...

Embodiment 2

[0047] Animal immune experiment verification

[0048] Select 2 adult healthy male New Zealand white rabbits. After the synthesized whole antigen is fully mixed with Freund's complete adjuvant, the rabbits are immunized by dot injection on the neck and back. After 7 times of booster immunization, the test tube method and agar The antiserum titers of the double-diffusion test have reached 1:320 (1:32) and 1:640 (1:64), respectively, and the agar double-diffusion test shows that the cross-reaction is not obvious, indicating that its specificity is better.

Embodiment 3

[0050] Determination of PBBs by Direct Competitive Real-time Immuno-PCR

[0051] Take a clean 0.2mL polypropylene PCR tube and treat with 50μL of 0.8% glutaraldehyde solution at 37°C for 5-6h. Wash with ultrapure water for 3 min×3 times. Dilute the PBB-OVA solution with coating buffer (CBS), add it to a small PCR tube treated with glutaraldehyde (20 μL / well), overnight at 4°C, wash the tube with washing solution (PBST) for 3 min×3 times. Add 200 μL of PBS solution containing 3% OVA to each tube, incubate at 37° C. for 1 h, wash the tube with PBST for 3 min×3 times. Add 25 μL of PBB28 small molecule and 25 μL of biotinylated anti-PBB28 polyclonal antibody to the same PCR tube, incubate at 37°C for 1 h, and wash the tube with PBST for 3 min×3 times. Add 50 μL of 6.5 μg / mL avidin solution to each tube, and incubate at 37° C. for 0.5 h. Wash three times with PBST, each 3min. Add 50 μL of biotinylated DNA to each tube, incubate at 37°C for 1 hour, wash with PBST for 5 times, th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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 [0001] The invention relates to an immunogen and a preparation method thereof, in particular to a polybrominated biphenyl homologue immunogen and a preparation method thereof. Background technique [0002] Brominated flame retardants are currently one of the largest organic flame retardants in the world, mainly polybrominated diphenyl ethers (PBDEs) and polybrominated biphenyls (PBBs), which are often added to resins, polystyrene and polyurethane foams as flame retardant additives Among polymer synthetic materials such as polymers, it is widely used in plastic products, textiles, circuit boards and building materials, and slowly dissipates into the environment with the use of these products. [0003] The chemical properties of polybrominated biphenyls and polychlorinated biphenyls are similar, and they are very stable in the environment and easily form bioaccumulative chemicals. The understanding of the biological toxicity of polybrominated biphenyls has also...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765
Inventor 庄惠生陈寒玉
Owner SHANGHAI JIAO TONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap