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Troponin I autoantibody detection method based on gold nanoplates

An autoantibody and detection method technology, which is applied in the application field of nano-biotechnology to achieve the effects of high sensitivity, good stability and repeatability, and easy operation

Inactive Publication Date: 2012-05-02
JIANGSU PROVINCE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether cTnl autoantibodies in the serum of patients with coronary heart disease have a pathological effect on myocardial injury and whether it is related to the prognosis of patients remains to be further studied.

Method used

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  • Troponin I autoantibody detection method based on gold nanoplates
  • Troponin I autoantibody detection method based on gold nanoplates
  • Troponin I autoantibody detection method based on gold nanoplates

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Experimental program
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Embodiment 1

[0023] Embodiment 1 Preparation and acquisition of high-purity gold nanosheets

[0024] The preparation of gold nanosheets is based on the iodide ion (I - ) and trimethylcetylammonium bromide (CTAB) in the presence of "seed" growth strategy, first 1mL 10mM sodium citrate was added to 37.6mL of water, then 0.4mL 25mM chloroauric acid ( HAuCl 4 ) solution, and finally add 1 mL of 0.1M sodium borohydride (NaHB 4 ) solution. The color of the reaction solution immediately changed from light yellow to orange, indicating the formation of gold nanoparticles, which were used as crystal seeds. A typical procedure for generating gold nanosheets is as follows: 100 mL containing 0.25 mM HAuCl 4 and 0.05M CTAB growth solution were placed in a 250mL flask, and 55μL of 0.1M potassium iodide (KI), 0.55mL of 0.1M L-ascorbic acid (AA) and 0.5mL of 0.1M NaOH (NaOH The role here is to deprotonate AA for more efficient reducing power). Finally, 0.1 mL of "seed" was added to the growth solutio...

Embodiment 2

[0025] The gold nanosheet prepared in embodiment 2 is characterized by electron microscopy

[0026] The electron microscope observation results of angular gold nanosheets and spherical gold nanoparticles are as follows: figure 2 As shown, after separation of the mixture of triangular gold nanosheets and spherical gold nanoparticles, both have a narrow size distribution. The average diameter of the spherical gold nanoparticles is 30±2 nm. The average side length of the triangular gold nanosheets is 140±25nm, and the thickness is 8±2nm. Statistics from the TEM photos of the gold nanosheets show that the number of gold nanosheets accounts for more than 97% of the total number of particles. The basal plane of a single gold nanosheet was characterized by high-resolution electron microscopy ( figure 2 c), the lattice textures with the same orientation and the same spacing show that these gold nanosheets have the properties of single crystals. The electron diffraction pattern of...

Embodiment 3c

[0027] Example 3 cTnI antigen coated gold nanosheets, spherical gold nanoparticles

[0028] The resuspension of the gold nanosheets was centrifuged to remove free CTAB molecules in the solution, and the CTAB bilayer on the surface of the gold nanosheets was surface-modified with polystyrene sodium sulfonate (PSS) before binding to the antigen. The operation steps are as follows: 2.4 mL of 2.5 mg / mL PSS solution was added to 30 mL of the gold nanosheet solution whose optical intensity of the long-wave SPR peak was 1.0 under stirring, and then stood still for 15 minutes. Free PSS molecules in the solution system were removed by centrifugation. The pellet was resuspended with the same volume of deionized water and washed with 0.2M K 2 CO 3The pH of the solution was adjusted to 8.2. Then the PSS-coated gold nanosheet solution was reacted with excess cTnI antigen (50 μg / mL) for 30 min. Afterwards, 1% bovine serum albumin (BSA) in Tris buffer was added to block the unbound sites....

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Abstract

The invention belongs to the field of the nanometer biological technology application, and discloses a troponin I autoantibody detection method based on gold nanoplates. The method includes the following steps: coating a cardiac troponin I antigen on the gold nanoplate, detecting gold nanoplates with a spectrometer using specific binding between the cardiac troponin I antigen coated on the gold nanoplate and a cardiac troponin I autoantibody in a sample to be measured, thereby realizing the quantitative determination of the cardiac troponin I autoantibody. The detection method has the advantages of good stability, repeatability and simple operation, and a new approach of gold nanoplate is developed in the field of biomedical science.

Description

technical field [0001] The invention belongs to the field of nanobiological technology application, and relates to a method for detecting troponin I autoantibodies based on gold nanosheets. Background technique [0002] Gold nanoparticles have optical properties based on surface plasmon resonance (SPR), which originates from the resonant coupling between incident light of a specific wavelength and free electrons on the surface of gold nanoparticles. When the aggregation state of gold nanoparticles changes or the surface is adsorbed by different chemical molecules or biomolecules, its SPR peak will move or the peak shape will change. (UV-vis) spectrometer can be carried out. Theoretical and experimental studies in recent years have shown that the SPR peaks of asymmetrically shaped gold nanoparticles are more sensitive than those of spherical gold nanoparticles. However, there are still many challenges in using asymmetrically shaped gold nanoparticles as a biological detecti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33G01N1/34
Inventor 杨笛陈相健顾春荣郭志睿卞智萍吴恒芳徐晋妉
Owner JIANGSU PROVINCE HOSPITAL
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