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Preparation method of poly(sulfydryl)-modified composite nanoparticle probe of covalent coupling antibody

A technology of composite nanoparticles and covalent coupling, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of false positive results, instability, agglomeration and oxidation of immunodiagnostic results, and eliminate the influence of steric hindrance, which is not easy to detachment effect

Inactive Publication Date: 2018-06-29
NANJING TZONE BIOLOGICAL SCI & TECH
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  • Abstract
  • Description
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Problems solved by technology

These nanoparticles are widely used in the fields of bioanalysis and biomedicine, but they also have some limitations. The specific surface area of ​​magnetic nanoparticles is large, and their surface activity is extremely high, so they are prone to agglomeration and oxidation. The complex is unstable in the process of immunochromatography and may be separated, which may lead to false positive results of immunodiagnosis and other erroneous results

Method used

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  • Preparation method of poly(sulfydryl)-modified composite nanoparticle probe of covalent coupling antibody

Examples

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Effect test

Embodiment 1

[0021] Fc of covalently conjugated antibodies modified with mercaptooctadecanoic acid and mercapto-polyethylene glycol-carboxylates 2 o 3 Preparation of Au / Au composite nanoparticle probes:

[0022] 1) To prepare magnetic nanoparticles, FeCl 3 ·6H 2 O and FeSO 4 ·7H 2 O was added to the deoxidized hydrochloric acid solution to dissolve evenly. During the dissolution process, nitrogen protection was introduced to prevent oxidation. Add ammonia water at 25°C, stir rapidly for 20min, then heat the solution to 60°C in a water bath, and keep for 1 ~2h, collect magnetic nanoparticles with a magnet, wash with water several times, and store in water at low temperature.

[0023] 2) To prepare gold nanoparticles, pour 200mL of water into a 500mL beaker, then add 2mL of HAuCl 4 4H 2 O solution, place it on a magnetic constant temperature stirrer and heat it. When the liquid reaches a sub-boiling state (the temperature reaches about 98°C), quickly add 4mL sodium citrate solution to...

Embodiment 2

[0027] Fc of covalently conjugated antibodies modified with mercaptooctadecanoic acid and mercapto-polyethylene glycol-carboxylates 2 o 3 / Au / Ag composite nanoparticle probe preparation:

[0028] Using the same Fe as in Example 1 2 o 3 / Au composite nanoparticle preparation method, the prepared Fe 2 o 3 / Au composite nanoparticles were dispersed in water, and 0.7 mL of AgNO was slowly added while stirring 3 and sodium citrate mixed solution, and at the same time add HCHO solution and NH 3 ·H 2 O adjusts the pH of the solution to 10 and forms Fe 2 o 3 / Au / Ag composite nanoparticles, the product is magnetically separated, washed with water several times and then dried to constant weight.

[0029] to Fe 2 o 3 / Au / Ag composite nanoparticles were modified with polysulfhydryl groups, and the blank group was set as unmodified Fe 2 o 3 / Au / Ag composite nanoparticles, the test group was Fe modified by a mixture of mercaptooctadecanoic acid and mercapto-polyethylene glycol-...

Embodiment 3

[0031] Fc of covalently conjugated antibody modified by cystine and mercaptooctadecanoic acid 2 o 3 / Au / Ag composite nanoparticle probe preparation:

[0032] Using the same Fe as in Example 2 2 o 3 / Au / Ag composite nanoparticle preparation method and the same modification coupling condition steps, set the blank group as unmodified Fe 2 o 3 / Au / Ag composite nanoparticles, set the experimental group cystine and mercaptooctadecanoic acid mixed with Fe modified by 3:2 2 o 3 / Au / Ag composite nanoparticle, after the functional group modification and activation is successful, it is coupled with antibody protein to form a probe.

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Abstract

The invention relates to a preparation method of a poly(sulfydryl)-modified composite nanoparticle probe of a covalent coupling antibody. The preparation method is characterized by modifying the surfaces of gold nanoparticles, silicon dioxide nanoparticules, silver nanoparticles, magnetic nanoparticles or two composite nanoparticles by using a poly(sulfydryl) compound, thus forming functional groups with an amino group, a carboxyl group, a hydroxyl group and the like on the surface; covalently combining the functional group with an antibody through a coupling agent, thus forming a stable nanoprobe which is used for target to-be-detected object detection in an in-vitro diagnostic reagent or disease diagnosis. The poly(sulfydryl)-modified composite nanoparticle probe of the covalent coupling antibody, prepared through the preparation method, has many advantages of having stable performance and strong combining capacity with protein, greatly removing steric hindrance influence, and the like, has a good application prospect in fields of immunologic diagnosis, molecular diagnosis and the like, and has a very good application prospect in all fields of biomedicine.

Description

technical field [0001] The invention relates to a preparation method of a polythiol-modified covalently coupled antibody composite nanoparticle probe, which belongs to the field of in vitro diagnosis. Background technique [0002] Magnetic nanoparticles and noble metal nanoparticles are new functional materials. MNPs have the characteristics of small particle size, large specific surface area, easy surface modification, biocompatibility, low toxicity, etc., and also have unique magnetic responsiveness, which can be aggregated and positioned under a constant magnetic field. Noble metal nanoparticles, commonly gold nanoparticles and silver nanoparticles, have unique optical properties, easy surface modification, strong reducibility, high chemical stability, and good biocompatibility. . These nanoparticles are widely used in the fields of bioanalysis and biomedicine, but they also have some limitations. The specific surface area of ​​magnetic nanoparticles is large, and their...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/54346
Inventor 徐林
Owner NANJING TZONE BIOLOGICAL SCI & TECH
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