Synthesis method of fluorescent reagent for specific recognition and sensitive detection of human serum protein

A technology of human serum albumin and fluorescent reagents, applied in biological testing, fluorescence/phosphorescence, measuring devices, etc., can solve the problems of complex reagents, expensive instruments, unfavorable daily detection, etc.

Active Publication Date: 2019-02-05
宜都市华阳化工有限责任公司
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Problems solved by technology

At present, there are endless immunochemical methods for albumin detection, but most of them are not conducive to daily detection due to the complicated procedures and expensive reagents and instruments.

Method used

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  • Synthesis method of fluorescent reagent for specific recognition and sensitive detection of human serum protein
  • Synthesis method of fluorescent reagent for specific recognition and sensitive detection of human serum protein
  • Synthesis method of fluorescent reagent for specific recognition and sensitive detection of human serum protein

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Embodiment Construction

[0025] The following experiments and examples serve to further illustrate but not limit the present invention.

[0026] (1) In PBS medium, DB-15C5 is used to identify HSA and BSA, the method is as follows:

[0027] Weigh DB-15C5 0.01345g, accurately add 5mL of absolute ethanol to obtain 5.0×10 -3 mol / L mother liquor. Dilute the mother liquor with ethanol to 1.0 × 10 -4 The mol / L solution is ready for use, wrapped in plastic wrap and tinfoil, sealed, protected from light, and stored in the refrigerator. At the same time, human serum albumin (HSA) was prepared in phosphate buffer to 1.0×10 -4 mol / L solution for later use.

[0028] Take 10 μL of DB-15C5 mother solution in a quartz fluorescence cuvette, and then add 1 mL of phosphate buffer saline (PBS, pH 7.2 to 7.4) to obtain 5.0×10 -5 mol / L of the test solution. Add 200 μL of protein stock solution, and set 360 nm as the excitation wavelength on the fluorescence spectrophotometer. The excitation and emission slits were t...

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Abstract

The invention belongs to the field of biological macromolecular probes, and particularly relates to a fluorescent reagent and a method for measuring human albumin and efficiently and quickly identifying the human albumin and bovine serum albumin. A schiff base organic compound with a typical torsion intramolecular charge transfer feature is designed and synthesized on the basis of a triphenylamine and 15-crown-5 ester structure, and is used as the fluorescent reagent; protein is added into a neutral phosphate buffer solution, the human albumin can be quickly identified from several types of common proteins according to compound fluorescent spectrum features, and a standard curve is established to quantitatively measure the contents of serum proteins in a buffered saline solution and artificial urine. The concentration of the probe reagent is 50mu M, and the 5mu M human albumin can be identified; after quantitative detection, the range of the human albumin in the buffered saline solution is 0 to 5.6mu M, and the minimum detection limit is 0.112mg / L; the range of the human albumin in the artificial urine is 0 to 1.2mu M, and the minimum detection limit is 1.95mg / L.

Description

technical field [0001] The invention relates to a fluorescent probe method for measuring human serum protein concentration and a method for identifying it and bovine serum protein, belonging to the field of biomacromolecule content and structure research. Background technique [0002] In clinical diagnosis, identification of human serum protein in urine is an early signal of kidney disease, and insufficient protein content in human serum can also indicate liver failure. In addition, low-cost bovine serum albumin (BSA) is widely used to replace human serum albumin (HSA) in biochemical or pharmacological applications. However, only 75.8% of the biological functions of bovine serum albumin are similar to human serum albumin, and it cannot replace human serum albumin to replace fluid and help surgical patients restore blood volume. Therefore, it is particularly important to distinguish between human serum albumin and bovine serum albumin during treatment, and to achieve quantit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N33/68
CPCG01N21/64G01N33/68G01N2333/46G01N2333/765
Inventor 杨昌英沈萍金红都华静怡杨威刘春琳
Owner 宜都市华阳化工有限责任公司
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