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Method for quantitatively assaying 80nmAuNPs on the basis of indirect competitive enzyme-linked immunosorbent assay

An enzyme-linked immunosorbent assay technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of time-consuming, complicated operation, and complicated operation, and achieve the effect of low detection limit, simple operation and high sensitivity.

Active Publication Date: 2015-12-23
重庆陌达生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Both methods have certain limitations. The first method is limited to the measurement of metal nanoparticles, and has great limitations for the quantitative detection of non-metal nanoparticles. When it is prepared into a solution, it is necessary to find a specific solute that can dissolve the nanomaterial to be tested. Secondly, the solute must be subjected to ultraviolet spectrophotometry to establish the relationship between its wavelength and absorbance. Finally, the measurement wavelength of the nanomaterial to be tested is determined by a comparison method. The method is more complicated, and the operation is relatively cumbersome; the pretreatment of the sample in the second method is more cumbersome, the operation is complicated, time-consuming, and the experimental equipment used is relatively expensive

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  • Method for quantitatively assaying 80nmAuNPs on the basis of indirect competitive enzyme-linked immunosorbent assay
  • Method for quantitatively assaying 80nmAuNPs on the basis of indirect competitive enzyme-linked immunosorbent assay

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Embodiment 1

[0059] A method for quantitatively detecting 80nmAuNPs based on an indirect competitive enzyme-linked immunosorbent assay, comprising the following steps:

[0060] (1) Synthesis of 80nm AuNPs whole antigen

[0061] 1) Preparation of 80 nm AuNPs

[0062] Take 500 μL of 2% wtHAuCl 4 and 99mL of distilled water in a three-necked flask, heated to boiling with a collector-type constant temperature heating magnetic stirrer, the heating to the boiling process took 15min-30min, and then quickly added 0.7mL of 1% trisodium citrate to the boiling solution, and continued to boil for 35min. , removed the heating device, stirred and cooled to room temperature, collected and stored at 4°C in the dark, and this earthy red solution was 80 nm AuNPs.

[0063] 2) Preparation of 80nm AuNPs-coated antigens and immunogens

[0064] Take 8 mL of the above 80 nm AuNPs solution, add 20 μL of 9.8% thioglycolic acid (TGA) under stirring for 15 min, and then continue to add 10 μL of 0.09 mol / L hydroxys...

Embodiment 2

[0084] A method for quantitatively detecting 80nmAuNPs based on an indirect competitive enzyme-linked immunosorbent assay, comprising the following steps:

[0085] (1) Synthesis of 80nm AuNPs whole antigen

[0086] 1) Preparation of 80 nm AuNPs

[0087] Take 500 μL of 2wt% HAuCl 4 and 99mL of distilled water in a three-necked flask, heated to boiling with a collector-type constant temperature heating magnetic stirrer, and the heating to boiling process took 15min-30min, and then quickly added 0.7mL of 1wt% trisodium citrate to the boiling solution, and continued to boil for 35min. , removed the heating device, stirred and cooled to room temperature, collected and stored at 4°C in the dark, and this earthy red solution was 80 nm AuNPs.

[0088] 2) Preparation of 80nm AuNPs-coated antigens and immunogens

[0089] Take 8 mL of the above 80 nm AuNPs solution, add 20 μL of 9.8% thioglycolic acid (TGA) under stirring for 13 min, and then continue to add 10 μL of 0.09 mol / L hydrox...

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Abstract

The invention provides a method for quantitatively assaying 80nmAuNPs on the basis of indirect competitive enzyme-linked immunosorbent assay, which adopts indirect competitive enzyme-linked immunosorbent assay in the technology of enzyme-linked immunosorbent assay and utilizes the specific binding of antigens and antibodies to assay 80nmAu particles. Compared with the prior art, the method successfully synthesizes haptens into holoantigens by the action of modifier, making great contribution to the production of antibodies. By utilizing the immune response effect of the body, the method successfully prepares an anti-80nmAuNPs polyclonal antibody with high specificity, the shelf life of the antibody is long, and therefore feasibility is high; by utilizing indirect competitive enzyme-linked immunosorbent assay in the technology of enzyme-linked immunosorbent assay to carry out a type of quantitative assay, the invention of the method provides a highly feasible method for the quantitative assay of future nanomaterials. The method is easy to operate, highly feasible, highly sensitive and low in assay limit, and can realize high-throughput assay.

Description

technical field [0001] The invention relates to the quantitative detection of nanomaterials, in particular to a method for quantitatively detecting 80 nm AuNPs based on an indirect competitive enzyme-linked immunosorbent assay. Background technique [0002] The development of society is bound to be accompanied by the progress of science and technology. With the advent of nanotechnology at the end of the 20th century, nanomaterials is a new and rapidly developing material science. Nanomaterials refer to ultrafine materials with particle size in the nanometer scale (1nm to 100nm), which have small size effect, large specific surface area, quantum size effect and macroscopic quantum tunneling effect. It is precisely because of these special properties that nanomaterials have been more and more widely used in life and production. For example, nanomaterials have very important applications in biosensors, bioprobes, drug carriers, medical imaging, sunscreens, etc. . However, tec...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/543
Inventor 张明翠张月
Owner 重庆陌达生物科技有限公司
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