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Synthesis method of fluorescent reagent for identifying specificity and sensitively detecting human albumin and application

A human serum protein, sensitive detection technology, used in biological testing, fluorescence/phosphorescence, measurement devices, etc.

Active Publication Date: 2017-05-10
宜都市华阳化工有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 identifying specificity and sensitively detecting human albumin and application
  • Synthesis method of fluorescent reagent for identifying specificity and sensitively detecting human albumin and application
  • Synthesis method of fluorescent reagent for identifying specificity and sensitively detecting human albumin and application

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Embodiment Construction

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

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

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

[0028] Take 10μL of DB-15C5 mother solution in a quartz fluorescent cuvette, and add 1mL phosphate buffer (PBS, pH 7.2~7.4) to obtain 5.0×10 -5 mol / L of the test solution. Add 200μL of protein stock solution, and set 360nm as the excitation wavelength on the fluorescence spectrophotometer. The excitation and emission slits are adjusted to 2.5, 5nm. Sc...

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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 the concentration of human serum protein and identifying it with bovine serum protein, which belongs to the research field of biological macromolecule content and structure. Background technique [0002] In clinical diagnosis, the identification and identification of human serum protein in urine is an early signal of kidney disease, and the lack of 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 the fields of biochemistry or pharmacology. However, only 75.8% of the biological functions of bovine serum albumin are similar to those of human serum albumin, and it cannot replace human serum albumin to replace fluids and help surgical patients repair blood volume. Therefore, it is particularly important to distinguish between human serum protein and bovine serum protein during t...

Claims

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Application Information

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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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