Sarcosine detection method based on metal organic framework material

A metal-organic framework and detection method technology, applied in the field of chemical detection, can solve the problems of reducing the catalytic efficiency of cascade reactions and achieve the effect of increasing the concentration of substrates

Active Publication Date: 2020-01-17
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, SOX and HRP are in a disordered state in solution, and random substrate collisions significantly reduce the catalytic efficiency of the cascade reaction.

Method used

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  • Sarcosine detection method based on metal organic framework material

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Effect test

Embodiment 1

[0020] A metal-organic framework material-based sarcosine detection method, based on the metal-organic framework material wrapping sarcosine oxidase and catalase, thereby promoting the pair of cascade enzymes to be confined in a small space, through the cascade reaction Adopt the method for chemiluminescence to measure the content of sarcosine afterward, follow the steps below:

[0021] (1) ZIF-8 wraps SOX and HRP:

[0022] Weigh 0.1 mg of SOX and 0.2 mg of HRP and dissolve in 1 mL of 1 M 2-methylimidazole (2-MIM) aqueous solution, mix well at room temperature and incubate for 10 minutes to obtain a mixture; then, in the mixture Quickly add 1 mL of 20 mM zinc acetate solution, mix well, and mechanically stir overnight at room temperature. The mixture was centrifuged at 5000 rpm / min for 15 min, washed twice with ethanol, and the precipitate was respun with a solution containing 2% PVP (w / w), mechanically stirred for 1 hr; then, centrifuged at 5000 rpm / min for 20 min, collect ...

Embodiment 2

[0026] A method for detecting sarcosine based on a metal organic framework material, similar to Example 1, according to the following steps:

[0027] (1) ZIF-8 wraps SOX and HRP:

[0028] Weigh 0.1 mg of SOX and 0.3 mg of HRP and dissolve in 1 mL of 1 M 2-methylimidazole (2-MIM) aqueous solution, mix well at room temperature and incubate for 10 minutes to obtain a mixture; then, in the mixture Quickly add 1 mL of zinc acetate solution with a concentration of 20 mM, mix well, and mechanically stir overnight at room temperature; the precipitate obtained by centrifuging the mixture at 5000 rpm / min for 15 min is washed twice with ethanol, and is washed with 2% PVP (w / w) The solution was re-spinned and centrifuged for 1 hr; then, centrifuged at 5000 rpm / min for 20 min to collect PVP-stabilized SOX-HRP@ZIF-8, and the precipitate was dispersed in 2 mL of phosphate buffer ( PBS, pH 7.4) to get the PVP-SOX-HRP@ZIF-8 solution.

[0029] (2) Determination of sarcosine content:

[0030...

Embodiment 3

[0032] A method for detecting sarcosine based on a metal organic framework material, similar to Example 1, according to the following steps:

[0033] (1) ZIF-8 wraps SOX and HRP:

[0034] Weigh 0.2 mg of SOX and 0.3 mg of HRP and dissolve in 1 mL of 1 M 2-methylimidazole (2-MIM) aqueous solution, mix well at room temperature and incubate for 10 minutes to obtain a mixture; then, in the mixture Quickly add 1 mL of zinc acetate solution with a concentration of 20 mM, mix well, and mechanically stir overnight at room temperature; / w) The solution was re-spinned and centrifuged for 1 hr; then, centrifuged at 5000 rpm / min for 20 min to collect PVP-stabilized SOX-HRP@ZIF-8, and the precipitate was dispersed in 2 mL of phosphate buffer (PBS, pH 7.4) to get PVP-SOX-HRP@ZIF-8 solution;

[0035] (2) Determination of sarcosine content:

[0036] Use PBS to prepare 10 mM ABTS solution, prepare 5-100 µM sarcosine solution, and store in ice bath; turn on the UV spectrophotometer, set the ...

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Abstract

The invention relates to a sarcosine detection method based on a metal organic framework material. The sarcosine detection method is characterized in that sarcosine oxidase and catalase are wrapped bymeans of the metal organic framework material, so that the pair of cascade enzymes is limited in a smaller space, and the content of sarcosine is determined by adopting a chemiluminescence method after a cascade reaction. The sarcosine detection method adopts ZIF-8 to coat SOX and HRP; ZIF-8 has a large specific surface area and porous channels, so that a reaction substrate in a solution can penetrate through the channels to react with SOX and HRP, enrichment of the substrate around the enzymes is facilitated, and the local substrate concentration is increased; meanwhile, due to wrapping of ZIF-8, SOX and HRP are limited within the range of hundreds of nanometers, the distance between SOX and HRP is regulated and controlled, hydrogen peroxide generated by catalyzing sarcosine by means ofSOX can be effectively transmitted to HRP for a chromogenic reaction, and the reaction rate of the reaction cascade enzymes is increased.

Description

technical field [0001] The invention relates to a detection method for detecting sarcosine, a specific target of prostate cancer, in particular to a detection method for sarcosine based on a metal organic framework material. The invention belongs to the field of chemical detection. Background technique [0002] Prostate cancer is one of the common male malignant tumors. Especially with the aging of Chinese society, the diagnosis rate and death rate of prostate cancer will increase. Therefore, early screening of prostate cancer patients can significantly improve and improve the survival of patients Rate. Existing diagnostic methods for prostate cancer are mainly based on prostate-specific antigen (PSA), which has serious problems in detection sensitivity and specificity, and has a high false positive rate and false negative rate, so many patients undergo unnecessary biopsies. caused great pain. Therefore, a new marker detection method and an effective screening method are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N21/78G01N21/31
CPCG01N21/76G01N21/78G01N21/31
Inventor 崔大祥张兆坤徐艳陈玮嘉朱君金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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