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Novel synthesis method of polyfluorene derivative and application in immunosensor for detecting tumor marker

A derivative and fluorescence sensor technology, applied in the field of immune marker synthesis and immune detection, can solve the problems of unfavorable popularization, long time consumption, complicated operation, etc., and achieve the effect of reducing impact, improving biocompatibility, and high fluorescence efficiency

Inactive Publication Date: 2013-04-10
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional immunoassay techniques such as ELISA, radioimmunoassay, and colloidal gold immunoassay generally take a long time and require cumbersome operations such as adding samples, incubating, and washing, while the new immunoassay method high-performance liquid chromatography chemiluminescence immunoassay , microfluidic chip chemiluminescence immunoassay, capillary electrophoresis chemiluminescence immunoassay, etc. generally require expensive instruments, which is not conducive to popularization

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  • Novel synthesis method of polyfluorene derivative and application in immunosensor for detecting tumor marker
  • Novel synthesis method of polyfluorene derivative and application in immunosensor for detecting tumor marker
  • Novel synthesis method of polyfluorene derivative and application in immunosensor for detecting tumor marker

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preparation example Construction

[0043] A method for preparing a flow injection fluorescent sensor for detecting immune markers, comprising the following steps:

[0044] (1) Synthesis of polyfluorene derivatives (PF-NH 2 );

[0045] (2) Synthesis of silica-coated polyfluorene derivatives (PF-NH 2 SiO 2 );

[0046] (3) Modify the secondary antibody in PF-NH 2 SiO 2 ;

[0047] (4) Prepare gold nanoparticles (AuNPs), ferroferromagnetic particles (Fe 3 o 4 ) Gold nanoparticles coated iron oxide (Fe 3 o 4 Au);

[0048] (5) Modify the primary antibody on Fc 3 o 4 Au surface;

[0049] (6) Fabricate flow injection fluorescent flow cells using polymethyl methacrylate (PMMA);

[0050] (7) Combine the prepared chemiluminescence sensor with a flow injection chemiluminescence instrument to detect immune markers.

[0051] The preparation of the silicon dioxide-coated polyfluorene derivative and the silicon dioxide-coated ferric oxide of the present invention comprises the following steps:

[0052] (1) 1,4-...

Embodiment 1

[0071] Example 1 (Carbohydrate markers, such as CA19-9)

[0072] (1) First synthesize three monomers: 1,4-dibromo-p-phenylenediamine (compound 1), 2,7-dibromo-9,9-bis(3′-(N, N′-dimethylamino )Propyl)-fluorene (compound 2) and p-divinylbenzene (compound 3). Polymerization into PF-NH by Heck 2 Synthesis.

[0073] (2) The PF-NH prepared in (1) 2 Prepare solution (1 nM in DMF). Add 1 mL PF-NH to a 100 mL one-necked bottle 2 solution, 8 mL of ethanol, stirred to disperse evenly. Then 0.1 mL of ammonia solution (25%) was added, and 10 μL of TEOS solution was stirred for 30 min. Continue to add 10 μL of TEOS solution and stir for 30 min until the total amount of TEOS solution added is 50 μL. After the reaction, at 10000 r min -1 Centrifuge at a rotating speed, wash the precipitate with ethanol, and repeat three times. Dry in a vacuum oven at 50 °C for 5 h to obtain PF-NH 2 SiO 2 .

[0074] (3) Add the PF-NH prepared in (2) into a small 10 mL beaker 2 SiO 2 0.02 g, 1 ...

Embodiment 2

[0083] Example 2 (Hormones, such as HCG)

[0084] (1) First synthesize three monomers: 1,4-dibromo-p-phenylenediamine (compound 1), 2,7-dibromo-9,9-bis(3′-(N, N′-dimethylamino )Propyl)-fluorene (compound 2) and p-divinylbenzene (compound 3). Polymerization into PF-NH by Heck 2 Synthesis.

[0085] (2) The PF-NH prepared in (1) 2 Prepare solution (1 nM in DMF). Add 1 mL PF-NH to a 100 mL one-necked bottle 2 solution, 8 mL of ethanol, stirred to disperse evenly. Then 0.1 mL of ammonia solution (25%) was added, and 10 μL of TEOS solution was stirred for 30 min. Continue to add 10 μL of TEOS solution and stir for 30 min until the total amount of TEOS solution added is 50 μL. After the reaction, at 10000 r min -1 Centrifuge at a rotating speed, wash the precipitate with ethanol, and repeat three times. Dry in a vacuum oven at 50 °C for 5 h to obtain PF-NH 2 SiO 2 .

[0086] (3) Add the PF-NH prepared in (2) into a small 10 mL beaker 2 SiO 2 0.02 g, 1 mL of CDI solut...

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Abstract

The invention discloses a novel synthesis method of a polyfluorene derivative and an application in a flow injection fluorescent sensor for a tumor marker. The synthesis method of the polyfluorene derivative comprises the following steps that: 1,4-dibromo p-phenylenediamine (a compound 1) is synthesized; 2,7-dibromo-9,9-bi(3'-(N,N'-dimethylamino) propyl)-fluorene (a compound 2) is synthesized; p-vinyltoluene (a compound 3) is synthesized; the three monomers are polymerized into the polyfluorene derivative by a Heck reaction. A fabrication method of the sensor comprises the following steps that polymethyl methacrylate is adopted to fabricate a flow injection fluorescent flow cell; ferroferric oxide magnetic particles and gold nanoparticles are prepared and assembled into gold covered ferroferric oxide; a primary antibody is modified on gold cladded ferroferric oxide; the polyfluorene derivative is subjected to silicon cladding treatment; a secondary antibody is modified on the surface of the silicon cladded polyfluorene derivative; an immune reaction is conducted; and flow injection fluoroscopic detection is conducted on the tumor marker. The sensor is strong in specificity, high in sensitivity, simple to operate, and low in detecting line.

Description

[0001] The invention relates to the technical field of synthesis of immune markers and immune detection, more specifically to the synthesis cost of a novel polyfluorene derivative and a method for using the novel polyfluorene derivative as a flow injection fluorescent sensor for immune markers. Background technique [0002] Conjugated polymers are a class of polymers with long-range π-bond conjugated systems in structure. Since the development of conjugated polymers, there are many types, mainly polyphenylene (PPP), polyphenylene vinylene (PPV), polyethylene fast (PA), polysialene (PT), polyaniline (PANI), Polyphenylene sulfide (PPS), polypyrrole (PPY), polydiacetylene (PDA) and other conjugated polymers. Conjugated polymers are usually easy to interact with groups or substances with acceptor properties, so that the physical and chemical properties of these polymers themselves change, and some materials with specific functions are generated. The covalent bonds of the conjugat...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N21/64G01N21/76C08G61/02C09K11/06C09K11/02
Inventor 颜梅楚成超葛慎光葛磊于京华黄家栋李蒙李龙刘芳刘伟艳王衍虎张彦王盼盼李伟平
Owner UNIV OF JINAN
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