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A kind of insulin antibody imprinted polymer and its preparation method and application

A technique of imprinting polymers and insulin, which is applied in the field of bioengineering, can solve the problems of increased cost of Elisa kits, non-reusable use, and high cost of antigens, and achieve the effects of saving detection time, low cost, and high environmental tolerance

Active Publication Date: 2018-05-04
SINOCARE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the antigens of anti-insulin antibodies in the serum must be blocked by coating, which is time-consuming and laborious, and the use is cumbersome
Antigens are generally expensive and cannot be reused
Lead to higher cost of Elisa kit

Method used

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  • A kind of insulin antibody imprinted polymer and its preparation method and application
  • A kind of insulin antibody imprinted polymer and its preparation method and application
  • A kind of insulin antibody imprinted polymer and its preparation method and application

Examples

Experimental program
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preparation example Construction

[0034] Fabrication of Magnetically Imprinted Polymers: Preparation of Fe by Co-precipitation 3 o 4 nanoparticles; by sol-gel method in Fe 3 o 4 The surface of the nanoparticles is coated with a silicon shell to prepare a magnetic silicon sphere carrier. The magnetic bead carrier, acrylamide, methacrylic acid, template molecular protein, and cross-linking agent are added to the neutral PBS buffer and mixed, and the polymerization is initiated by the initiator. The polymer was eluted with deionized water, 36% NaCl solution by mass percentage, 10% SDS solution, and 10% acetic acid solution by volume to prepare the insulin antibody imprinted polymer. The mass ratio of the magnetic bead carrier to the template molecular protein is (55-65):11, and that of the template protein, acrylamide, and methacrylic acid is 1:(1-2):1.

[0035] The following is a description of the application of the insulin antibody imprinted polymer of the present invention in the ELISA detection method: ...

Embodiment 1

[0039] Example 1: Preparation of insulin antibody magnetic molecular imprinting material

[0040] Preparation of Magnetic Molecularly Imprinted Materials for Insulin Antibody: Preparation of Fe by Co-precipitation Method 3 o 4 Nanoparticles, 2.08g FeCl 2 4H 2 O, 3.2073g FeCl 3 Dissolve in 100mL sterile water, pass nitrogen gas, heat the solution to 70°C, add 12mL ammonia solution (mass fraction: 28%). Stir the reaction for 30 minutes. After the product is cooled, remove the supernatant with the help of an external magnetic field, and wash the black product with sterile water and ethanol several times to obtain superparamagnetic Fe 3 o 4 After drying the nanoparticles, 160 mL of ethanol and 40 mL of water were added, 8 mL of ammonia water (28% by mass) and 2 mL of TEOS were added. Stir the reaction for 10 h, wash with sterile water and ethanol after magnetic separation. dry.

[0041] Weigh 30mg insulin antibody, 50mg acrylamide, 30mg methacrylic acid, 10mg MBA, 300mg Fe...

Embodiment 2

[0042] Example 2: Preparation of insulin antibody magnetic molecular imprinting material

[0043] Preparation of Magnetic Molecularly Imprinted Materials for Insulin Antibody: Preparation of Fe by Co-precipitation Method 3 o 4 Nanoparticles, 2.08g FeCl 2 4H 2 O, 3.2073g FeCl 3 Dissolve in 100mL sterile water, pass nitrogen gas, heat the solution to 70°C, add 12mL ammonia solution (mass fraction: 28%). Stir the reaction for 30 minutes. After the product is cooled, remove the supernatant with the help of an external magnetic field, and wash the black product with sterile water and ethanol several times to obtain superparamagnetic Fe 3 o 4 After drying the nanoparticles, 160 mL of ethanol and 40 mL of water were added, 8 mL of ammonia water (28% by mass) and 2 mL of TEOS were added. Stir the reaction for 10 h, wash with sterile water and ethanol after magnetic separation. dry.

[0044] Weigh 40mg insulin antibody, 40mg acrylamide, 40mg methacrylic acid, 10mg MBA, 200mg Fe...

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Abstract

The invention relates to the field of bioengineering, in particular to an anti-insulin antibody imprinted polymer and a preparation method and application thereof. The imprinting technology and magnetic separation are combined and applied to ELISA (enzyme-linked immuno sorbent assay) testing of anti-insulin antibodies in serum. The anti-insulin antibody magnetic molecular imprinted polymer is of a core-shell structure taking a Fe3O4@SiO2 magnetic silicon ball carrier as a core and an anti-insulin antibody molecular imprinted polymer as a shell. Imprinted polymer microspheres replace conventional antigens, enveloping and closing processes are no longer performed, and testing time is saved greatly. The imprinted polymer microspheres are more stable than the conventional antigens, and are high in environmental tolerance, simple to prepare and store and low in cost. The used imprinted polymer microspheres can be used repeatedly, and are simple to elute. A 96 pore plate can be eluted and reused, and waste of materials is avoided. The anti-insulin antibody imprinted polymer is an environment-friendly product.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to an insulin antibody imprinted polymer and its preparation method and application. Background technique [0002] Insulin antibody is an important marker of type 1 diabetes. Insulin antibodies may be produced by the destruction of β cells, so the detection of insulin autoantibodies can be used as a sign of autoimmune β cell damage, which is of great significance for the diagnosis, differential diagnosis and treatment of diabetes and hypoglycemia. Therefore, the detection of anti-insulin antibodies in serum can help to detect diabetes in time. [0003] The existing detection of anti-insulin antibody in serum all uses kits for detection. This method is convenient and effective, and the kit specifically includes all reagents and control solutions for detecting anti-insulin antibodies in serum proteins, including carriers, antigens or antibodies, enzyme substrates, negative and positive...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F220/06C08F220/56C08F222/38C08K9/10C08K3/22C08J9/26G01N33/551G01N33/531G01N21/76
CPCC08F220/06C08F220/56C08J9/26C08J2333/02C08J2333/26C08K3/22C08K9/10C08K2003/2275C08K2201/011G01N21/76G01N33/531G01N33/551C08F222/385
Inventor 张小娜刘伟
Owner SINOCARE