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Method for preparing quantum dot marked catalase fluorescent probe

A technology of catalase and fluorescent probe, which is applied in the field of preparation of CdTe quantum dot-labeled catalase fluorescent probe, can solve the problems of small surface area of ​​active groups, difficult target substances, binding and other problems, and achieve rapid preparation process Simple, Versatile, Inexpensive Effects

Inactive Publication Date: 2011-08-31
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the ligands selected for the quantum dots synthesized in the water phase are mostly simple mercapto compounds, the active groups on the surface are relatively small and the surface area is small, and it is difficult to combine with the target substance, so the direct application of the quantum dots in the water phase has limitations.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A CdTe quantum dot-labeled catalase fluorescent probe, which includes CdTe quantum dots, catalase (CAT, SIGMA-ALORICH), N-hydroxysuccinimide (NHS, Shanghai Yanchang Biochemical Company), L- Glycine (Glycine, Lingfei Technology), the rest is Tris buffer solution.

[0025] In the CdTe quantum dots, cadmium chloride (CdCl 2 2.5H 2 O, Sinopharm Chemical Reagent Co., Ltd.), potassium telluride hydride (KTeH 4 ,) and thioglycolic acid (TGA, Japan) at a molar ratio of 2.5:1:3, respectively, and heated at 100°C for 10–20 min.

[0026] The pH value of the Tris buffer solution is 6.8-7.6.

[0027] A kind of preparation method of CdTe quantum dot label catalase fluorescent probe, its steps are as follows:

[0028] A, preparation potassium hydride telluride (KH 4 Te) solution: through potassium borohydride (KBH 4 ) to reduce tellurium powder (Te), the reaction equation is as follows:

[0029] 4KBH 4 + 2Te + 7H 2 O → 2KH 4 Te + K 2 B 4 o 7 +11H 2 ↑

[0030] Weigh re...

Embodiment 2

[0037] A CdTe quantum dot labeled catalase fluorescent probe, which includes CdTe quantum dots (self-made), catalase (CAT,), N-hydroxysuccinimide (NHS,), L-glycine (Glycine, ), and the rest is Tris buffer solution.

[0038] In the CdTe quantum dots, cadmium chloride (CdCl 2 2.5H 2 O,), potassium telluride hydride (KTeH 4 , self-made), the molar ratio of thioglycolic acid (TGA,) was 2.5:1:3, and heated at 100°C for 10-20min.

[0039] The pH value of the Tris buffer solution is 7.2.

[0040] Take cadmium chloride (CdCl 2 ), potassium hydride telluride (KH 4 The molar ratio of Te) and thioglycolic acid (TGA) is a mixed solution of 2.5:1:3 respectively, adjust the pH value of the solution to 11.0, pack 50ml in each tank into polyvinyl fluoride digestion tanks, and heat at 100°C for 20min Prepared CdTe quantum dots. Take 4mL prepared concentration of 1 mmol / L (as Cd 2+ For CdTe quantum dots, add 0.2mL0.5g / L N-hydroxysuccinimide (NHS) and shake at 37℃ for 0.5h, then add 0.5m...

Embodiment 3

[0043] Take cadmium chloride (CdCl 2 ), potassium hydride telluride (KH 4 The molar ratio of Te) and thioglycolic acid (TGA) is a mixed solution of 2.5:1:3 respectively, adjust the pH value of the solution to 11.4, pack in polytetrafluoroethylene digestion tanks with 50ml per tank, and heat at 100°C with microwave CdTe quantum dots were prepared in 15 minutes. Take 4mL prepared concentration of 1 mmol / L (as Cd 2+ For CdTe quantum dots, add 0.2mL0.5g / L N-hydroxysuccinimide (NHS) and shake at 30℃ for 0.5h, then add 0.5mL4×10 -10 mol / L catalase (CAT) solution, shake at 30°C for 45 minutes, then add 0.3 mL, 0.1 mol / L L-glycine (Glycine) to terminate cross-linking, and prepare CdTe quantum dots labeled peroxide Hydrogenase fluorescent probe, the prepared fluorescent probe has a certain enzyme activity, and the fluorescence intensity is greatly improved (50%).

[0044] Other implementation steps are the same as in Example 1.

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Abstract

The invention discloses a method for preparing a CdTe quantum dot marked catalase fluorescent probe, which comprises the following steps: A) preparing a potassium hydrogen telluride solution by reducing the tellurium powder with potassium borohydride; B) weighting cadmium chloride crystals, and dissolving the cadmium chloride crystals in high purity water and fixing the volume and to obtain a cadmium chloride solution; C) weighting mercaptoacetic acid, diluting the mercaptoacetic acid with high purity water and fixing the volume to obtain a mercaptoacetic acid solution; D) taking anaerobic high purity water, stirring and continuously introducing high-purity nitrogen, respectively adding the cadmium chloride solution and the mercaptoacetic acid solution, adding the sodium hydroxide solution for adjusting the pH value, and then adding the potassium hydrogen telluride solution; E) canning the mixed solution into polytetrafluoroethylene digestion jars, and keeping the mole ratio of cadmium chloride, potassium hydrogen telluride and mercaptoacetic acid; and F) adding CdTe quantum dots into N-hydroxy succinimide and shaking at constant temperature to finally obtain the quantum dot marked catalase fluorescent probe. The method has simple process and the technical parameters are easily controlled. The biocompatibility is excellent. The probe has double detection property and can be used for fluorescent detection and enzymatic activity detection.

Description

technical field [0001] The invention relates to the biological application of quantum dots, and more specifically relates to a preparation method of a CdTe quantum dot-labeled catalase fluorescent probe, which can be used to detect the effects of pollutants, poisons and enzymes. Background technique [0002] Quantum dots synthesized in the organic phase are not hydrophilic, so they cannot be directly applied to biological systems, and must be modified on the surface. The operation process is complicated and the conditions are high. Quantum dots prepared by aqueous phase synthesis can be directly used in biological probes, and more and more attention has been paid to their research, and the research on this aspect is progressing rapidly. However, since the ligands selected for the quantum dots synthesized in the water phase are mostly simple mercapto compounds, the active groups on the surface are relatively small and the surface area is small, and it is difficult to combine ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 周培疆陈驰何振宇史广宇
Owner WUHAN UNIV
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