Synthetic method and application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphoric acid

Abstract

The invention provides a synthesis method and application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphoric acid, carbohydrate D-glucose-6-sodium phosphate is combined with the manganese-doped zinc sulfide quantum dots by utilizing excellent biocompatibility and hydrophilicity of carbohydrate D-glucose-6-sodium phosphate and utilizing an affinity principle between phosphate groups and metal cations, so that the manganese-doped zinc sulfide quantum dots of the glucose-6-phosphoric acid are obtained. The manganese-doped zinc sulfide quantum dot of glucose-6-phosphoric acid, which is simple, convenient and safe in synthesis process, is prepared, and has ultrahigh specificity and selectivity on glycopeptide; magnetic ferroferric oxide is synthesized through a hydrothermal reaction, the surface of the magnetic ferroferric oxide is coated with a dopamine layer serving as a coupling connecting agent to be grafted with manganese-doped zinc sulfide quantum dots, the magnetic manganese-doped zinc sulfide quantum dots are obtained, the grafted manganese-doped zinc sulfide quantum dots serve as anchoring points to functionalize D-glucose-6-sodium phosphate, and the magnetic manganese-doped zinc sulfide quantum dots are obtained. According to the present invention, the hydrophilic property and the processability are improved, the strong responsiveness to the external magnetic field is provided, the solid-liquid separation process can be simplified, and the simultaneous separation and enrichment of glycopeptides and phosphopeptides can be achieved.

Description

technical field

[0001] The invention relates to the field of functionalized nanomaterials and nanotechnology, in particular to a synthesis method and application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate. Background technique

[0002] Diabetes is a metabolic disease characterized by hyperglycemia, and the biomarkers of diabetes are diverse. Among them, autoantibodies have been confirmed, but the timely diagnosis ability for the inherent heterogeneity of diabetes needs to be improved urgently. In the pathogenesis of diabetes, protein glycosylation plays a key role. As a kind of protein modification, glycosylation widely exists in human proteins. Protein glycosylation has pervasive effects on cellular processes such as intercellular signal transduction, immune regulation, and cell differentiation. Studies have shown that abnormal protein glycosylation is related to physiological abnormalities and pathology, and some glycosylated proteins have become...

Examples

Embodiment 1

[0045] The synthetic method of the manganese-doped zinc sulfide quantum dot of glucose-6-phosphate comprises the following steps:

[0046] S1: Dissolve 1.8 g of zinc sulfate heptahydrate and 0.1 g of manganese chloride tetrahydrate in 20 mL of deionized water; pour the resulting solution into a three-necked flask with a capacity of 250 mL, and stir for 30 min under nitrogen protection; Dissolve 1.5 g of sodium sulfide nonahydrate in 5 mL of deionized water, add it to a three-necked flask, and continue stirring for 30 minutes to obtain manganese-doped zinc sulfide quantum dots;

[0047] S2: Dissolve 500 mg of disodium glucose-6-phosphate in a mixed solution containing 75 mL of acetonitrile, 150 μL of trifluoroacetic acid and 50 mL of deionized water, add it to the three-necked flask in step S1, and stir at 25° C. for 6 h;

[0048] S3: The product obtained in step S2 was fully washed with deionized water and ethanol, and dried overnight at 50° C. in a vacuum dryer to obtain mang...

Embodiment 2

[0052] Application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate in the enrichment of glycopeptides in HRP:

[0053] (1) Preparation of sample: horseradish peroxidase (HRP) at 25mmol / LNH 4 HCO 3 Enzymolysis in solution at 37°C for 16 hours;

[0054] (2) Enrichment: 0.5 mg of glucose-6-phosphate manganese-doped zinc sulfide quantum dots dispersed into 10 μL of 1% volume ratio of trifluoroacetic acid, 9% volume ratio of acetonitrile and 90% volume ratio of deionized water Add 2 μL of the sample prepared in step (1) to the centrifuge tube, enrich at 37°C for 30 min; wash thoroughly with a buffer solution of 0.5% by volume of phosphoric acid, 14.5% by volume of deionized water and 85% by volume of acetonitrile and centrifuged three times; add 10 μL of 50% volume ratio of acetonitrile and 50% volume ratio of deionized water eluate, elute at 37°C for 30 minutes, and centrifuge to obtain the supernatant;

[0055] (3) Mass spectrometry: Take 1 μL of the supern...

Embodiment 3

[0059] Application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate in the enrichment of glycopeptides in mixed proteins:

[0060] (1) Sample preparation: bovine serum albumin (BSA) was reductively alkylated with dithiothreitol and iodoacetamide, and then enzymatically hydrolyzed at 37°C for 16 hours; horseradish peroxidase (HRP) was added in 25mM NH 4 HCO 3 Enzymatic hydrolysis at 37°C for 16 hours in the solution; add bovine serum albumin (BSA) and horseradish peroxidase (HRP) to acetonitrile containing 1% volume ratio of trifluoroacetic acid and 9% volume ratio at a molar ratio of 1000:1 and 90% by volume deionized water in a centrifuge tube;

[0061] (2) Enrichment: 0.5 mg of the nanomaterial obtained in Example 1 is dispersed into 100 μL of 1% by volume trifluoroacetic acid containing the glycopeptide of step (1), 9% by volume of acetonitrile and 90% by volume of deionized In a centrifuge tube of water, enrich at 37°C for 30 min; fully wash with a buf...

technical field

[0001] The invention relates to the field of functionalized nanomaterials and nanotechnology, in particular to a synthesis method and application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate. Background technique

[0002] Diabetes is a metabolic disease characterized by hyperglycemia, and the biomarkers of diabetes are diverse. Among them, autoantibodies have been confirmed, but the timely diagnosis ability for the inherent heterogeneity of diabetes needs to be improved urgently. In the pathogenesis of diabetes, protein glycosylation plays a key role. As a kind of protein modification, glycosylation widely exists in human proteins. Protein glycosylation has pervasive effects on cellular processes such as intercellular signal transduction, immune regulation, and cell differentiation. Studies have shown that abnormal protein glycosylation is related to physiological abnormalities and pathology, and some glycosylated proteins have become...

Examples

Embodiment 1

[0045] The synthetic method of the manganese-doped zinc sulfide quantum dot of glucose-6-phosphate comprises the following steps:

[0046] S1: Dissolve 1.8 g of zinc sulfate heptahydrate and 0.1 g of manganese chloride tetrahydrate in 20 mL of deionized water; pour the resulting solution into a three-necked flask with a capacity of 250 mL, and stir for 30 min under nitrogen protection; Dissolve 1.5 g of sodium sulfide nonahydrate in 5 mL of deionized water, add it to a three-necked flask, and continue stirring for 30 minutes to obtain manganese-doped zinc sulfide quantum dots;

[0047] S2: Dissolve 500 mg of disodium glucose-6-phosphate in a mixed solution containing 75 mL of acetonitrile, 150 μL of trifluoroacetic acid and 50 mL of deionized water, add it to the three-necked flask in step S1, and stir at 25° C. for 6 h;

[0048] S3: The product obtained in step S2 was fully washed with deionized water and ethanol, and dried overnight at 50° C. in a vacuum dryer to obtain mang...

Embodiment 2

[0052] Application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate in the enrichment of glycopeptides in HRP:

[0053] (1) Preparation of sample: horseradish peroxidase (HRP) at 25mmol / LNH 4 HCO 3 Enzymolysis in solution at 37°C for 16 hours;

[0054] (2) Enrichment: 0.5 mg of glucose-6-phosphate manganese-doped zinc sulfide quantum dots dispersed into 10 μL of 1% volume ratio of trifluoroacetic acid, 9% volume ratio of acetonitrile and 90% volume ratio of deionized water Add 2 μL of the sample prepared in step (1) to the centrifuge tube, enrich at 37°C for 30 min; wash thoroughly with a buffer solution of 0.5% by volume of phosphoric acid, 14.5% by volume of deionized water and 85% by volume of acetonitrile and centrifuged three times; add 10 μL of 50% volume ratio of acetonitrile and 50% volume ratio of deionized water eluate, elute at 37°C for 30 minutes, and centrifuge to obtain the supernatant;

[0055] (3) Mass spectrometry: Take 1 μL of the supern...

Embodiment 3

[0059] Application of manganese-doped zinc sulfide quantum dots of glucose-6-phosphate in the enrichment of glycopeptides in mixed proteins:

[0060] (1) Sample preparation: bovine serum albumin (BSA) was reductively alkylated with dithiothreitol and iodoacetamide, and then enzymatically hydrolyzed at 37°C for 16 hours; horseradish peroxidase (HRP) was added in 25mM NH 4 HCO 3 Enzymatic hydrolysis at 37°C for 16 hours in the solution; add bovine serum albumin (BSA) and horseradish peroxidase (HRP) to acetonitrile containing 1% volume ratio of trifluoroacetic acid and 9% volume ratio at a molar ratio of 1000:1 and 90% by volume deionized water in a centrifuge tube;

[0061] (2) Enrichment: 0.5 mg of the nanomaterial obtained in Example 1 is dispersed into 100 μL of 1% by volume trifluoroacetic acid containing the glycopeptide of step (1), 9% by volume of acetonitrile and 90% by volume of deionized In a centrifuge tube of water, enrich at 37°C for 30 min; fully wash with a buf...

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