Transition metal oxide nanoprobe, preparation method and application

A transition metal and nanoprobe technology, applied in the field of nanomaterials and biological analysis, can solve the problems of limited metal tag carrying capacity, lack of protein spatial distribution information, and destruction of the quantitative process.

Pending Publication Date: 2021-09-21
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
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Problems solved by technology

Most of the nanoprobes currently used for protein molecular imaging at the single-cell level are based on the principle of antibody-antigen immune effects, and can achieve imaging of up to 5-8 markers through signal selection, but are only suitable for Semi-quantitative analysis of highly expressed proteins
The secondary antibody tag based on inorganic metal element labeling can realize the expression analysis of dozens of protein molecules. This quantitative analysis method based on inorganic mass spectrometry has high quantitative sensitivity, but its destructive quantitative process leads to protein spatial distribution information. lack of
At the same time, although using a secondary antibody to recognize the primary antibody can achieve signal amplification and imaging, the amount of carrying the metal label on the secondary antibody is limited, and the price is relatively expensive, and the primary antibody has species selectivity

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  • Transition metal oxide nanoprobe, preparation method and application
  • Transition metal oxide nanoprobe, preparation method and application
  • Transition metal oxide nanoprobe, preparation method and application

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preparation example Construction

[0026] Such as Figure 5 Shown, also provide the preparation method of this transition metal oxide nanoprobe, it comprises the following steps:

[0027] (1) Dissolve a mixture of biotin, N-hydroxysuccinimide NHS and 1-ethyl-3-(3'dimethylaminopropyl) carbodiimide EDC in 2-(N-morpholine) In the ethanesulfonic acid MES buffer solution, stir vigorously at room temperature for a period of time under the condition of avoiding light to obtain the activated biotin solution; afterward, high-speed centrifugation and purification to obtain the activated biotin, and re-dissolve in DMSO to obtain the first solution;

[0028] (2) dissolving 5-50 g / L biological protein molecule solution in sodium bicarbonate buffer, and mixing to obtain the second solution;

[0029] (3) Mix the first solution and the second solution evenly, react for a period of time to obtain biotin protein, and then dialyze to obtain a purified biotin protein solution;

[0030] (4) After mixing the transition metal elem...

Embodiment 1

[0046] Preparation of iron oxide probe

[0047] A) Biotin, NHS and EDC were mixed in MES buffer solution at a molar ratio of 1:1:1, and stirred vigorously for 30 minutes at room temperature in the dark to obtain an activated biotin solution. Then centrifuge in a high-speed centrifuge at a speed of 9000 rpm for 5 minutes, discard the supernatant, and redissolve the precipitate in DMSO to obtain a reaction solution 1.

[0048] B) adding bovine serum albumin (hereinafter referred to as BSA) to a final concentration of 10 grams per liter into 0.1 moles per liter of sodium bicarbonate buffer solution with a pH of 9, which is referred to as reaction solution 2;

[0049] C) Mix the solution 1 obtained in the above step A with the solution 2 obtained in step B evenly, wherein the ratio of BSA molecules to activated biotin molecules is 1:10, react in the dark for 3 hours, and then use a dialysis bag with a molecular weight cut off of 10,000 Dialyzed for 3 days and then freeze-dried to...

Embodiment 2

[0055] Preparation of cobalt hydroxide probe

[0056] A) Add 2 ml of cobalt nitrate solution with a concentration of 50 mmol per ml to 10 ml of fibronectin solution with a concentration of 20 mg per ml, stir for 5 minutes to make it evenly mixed, then add 0.2 ml with a concentration of 2 moles per liter The sodium hydroxide solution was used to raise the final pH to above 10. After vigorous stirring for 2 hours, it was allowed to stand for 12 hours for aging, and then dialyzed for 36 hours and freeze-dried to obtain cobalt hydroxide nanoparticles mineralized by fibronectin.

[0057] B) Biotin, NHS and EDC were mixed in MES buffer solution at a molar ratio of 1:1:1.2, and vigorously stirred for 50 minutes at room temperature in the dark to obtain an activated biotin solution. Then centrifuge in a high-speed centrifuge at a speed of 7000 rpm for 5 minutes, discard the supernatant, and redissolve the pellet in DMSO.

[0058] C) the fibronectin mineralized cobalt hydroxide nanopa...

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Abstract

The invention discloses a transition metal oxide nanoprobe, a preparation method and application. The nanoprobe is low in cost, capable of conducting fluorescence imaging on specific protein and capable of being used for simultaneous imaging of multiple kinds of protein. The transition metal oxide nanoprobe comprises transition metal oxide nano-particles and modification molecules compounded on the surfaces of the transition metal oxide nano-particles, wherein the transition metal oxide comprises oxides or hydroxides of three elements of iron, cobalt and nickel, and the surface modification molecules comprise biological protein molecules, biotin molecules and fluorescein molecules.

Description

technical field [0001] The present invention relates to the technical field of nanomaterials and biological analysis, in particular to a transition metal oxide nanoprobe, a method for preparing the probe, and the application of the transition metal oxide nanoprobe in cell-specific protein imaging analysis . Background technique [0002] Nanoprobes are a class of nanomaterials based on their physical and chemical properties that can be used in a medical imaging instrument to assist in improving imaging quality. Most of the nanoprobes currently used for protein molecular imaging at the single-cell level are based on the principle of antibody-antigen immune effects, and can achieve imaging of up to 5-8 markers through signal selection, but are only suitable for Semi-quantitative analysis of highly expressed proteins. The secondary antibody tag based on inorganic metal element labeling can realize the expression analysis of dozens of protein molecules. This quantitative analys...

Claims

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Application Information

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IPC IPC(8): C09K11/60C09K11/02C09K11/06G01N21/64B82Y20/00B82Y40/00
CPCC09K11/602C09K11/025C09K11/02C09K11/06G01N21/6456G01N21/6428B82Y20/00B82Y40/00G01N2021/6439C09K2211/1088
Inventor 王亚玲陈春英赵宇亮鲁婧怡
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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