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Electrochemical immunoassay method based on Dopamine embedded liposome

An immunoassay method and electrochemical technology, which is applied in the field of electrochemical immunoassay of small monolayer liposomes, can solve the problems affecting the accuracy of small cell lung cancer, the sensitivity of enzyme-linked immunoassay is not high, and it is difficult to detect trace amounts of gastric secretion. Precursin releasing peptide and other issues

Inactive Publication Date: 2011-10-26
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the detection of progastrin-releasing peptide (ProGRP) by enzyme-linked immunosorbent assay has been in the clinical application stage at home and abroad, but the sensitivity of enzyme-linked immunosorbent assay is not high, and it is difficult to detect trace amounts of progastrin-releasing peptide (ProGRP) , affecting the accuracy of detection of small cell lung cancer (SCLC)

Method used

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  • Electrochemical immunoassay method based on Dopamine embedded liposome
  • Electrochemical immunoassay method based on Dopamine embedded liposome
  • Electrochemical immunoassay method based on Dopamine embedded liposome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1 prepares the small unilamellar liposome of embedding dopamine

[0073] Mix 1 mg of a mixture of dipalmitoylphosphatidylcholine (DPPC) and cephalin (PE) with a molar ratio of 3:1 and 10 mL of a chloroform-methanol mixed solution with a volume ratio of 6:1, and ultrasonically treat it to obtain W / O emulsion; remove the organic solvent in the W / O emulsion by rotary evaporation under reduced pressure, add 2mL of phosphate buffer solution containing 0.1g / mL dopamine, swell in a water bath at 40°C for 80min, and mix vigorously to prepare embedding Dopamine-embedded multilamellar liposome vesicles: the dopamine-embedded multilamellar liposome vesicles were ultrasonically processed and centrifuged at 8000 rpm for 15 min to prepare dopamine-embedded small unilamellar liposomes. The prepared small unilamellar liposomes embedding dopamine were dispersed in PBS solution and stored at 4°C.

Embodiment 2

[0074] Embodiment 2 prepares the small unilamellar liposome of embedding dopamine

[0075] Mix 1 mg of a mixture of dipalmitoylphosphatidylcholine (DPPC) and cephalin (PE) with a molar ratio of 3:1 and 10 mL of a chloroform-methanol mixed solution with a volume ratio of 6:1, and ultrasonically treat it to obtain W / O emulsion; remove the organic solvent in the W / O emulsion by rotary evaporation under reduced pressure, add 2mL of phosphate buffer solution containing 0.1g / mL dopamine, swell in a water bath at 50°C for 60min, and mix vigorously to prepare embedding Dopamine-embedded multilamellar liposome vesicles; the dopamine-embedded multilamellar liposome vesicles were ultrasonically processed and centrifuged at 8000 rpm for 10 min to prepare dopamine-embedded small unilamellar liposomes. The prepared small unilamellar liposomes embedding dopamine were dispersed in PBS solution and stored at 4°C.

Embodiment 3

[0076] Embodiment 3 prepares the small unilamellar liposome of embedding dopamine

[0077] Mix 1 mg of a mixture of dipalmitoylphosphatidylcholine (DPPC) and cephalin (PE) with a molar ratio of 3:1 and 10 mL of a chloroform-methanol mixed solution with a volume ratio of 6:1, and ultrasonically treat it to obtain W / O emulsion; remove the organic solvent in the W / O emulsion by rotary evaporation under reduced pressure, add 2mL of phosphate buffer solution containing 0.1g / mL dopamine, swell in a water bath at 60°C for 40min, and mix vigorously to prepare embedding Dopamine-embedded multilamellar liposome vesicles; the dopamine-embedded multilamellar liposome vesicles were ultrasonically processed and centrifuged at 8000 rpm for 20 min to prepare dopamine-embedded small unilamellar liposomes. The prepared small unilamellar liposomes embedding dopamine were dispersed in PBS solution and stored at 4°C.

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Abstract

The invention discloses an electrochemical immunoassay method based on Dopamine embedded small single-layer liposome. The method comprises the following steps: ProGRP antibodies are subjected to a specific immunization reaction with ProGRPs in a sample to be detected; a conjugate of the dopamine embedded small single-layer liposome and the ProGRP antibodies, which is added and is demulsified by a surfactant, releases electroactive molecules Dopamine; an MWNT (multi-walled carbon nanotube) modified GCE (glassy carbon electrode) is treated as a working electrode, and the concentration of the ProGRP in the sample to be detected is detected through the signal intensity of a linear scanning current-voltage peak current of Dopamine. The electrochemical immunoassay method provided by the invention has a linear response range of 50 to 1000 pg / ml and a low detection limit of 16 pg / ml, so the method has the advantages of good specificity and high sensitivity, and has an important meaning to the SCLC (small-cell lung cancer) diagnosis.

Description

technical field [0001] The invention relates to the field of electrochemical immunity, in particular to an electrochemical immunological detection method based on small monolayer liposomes embedding dopamine. Background technique [0002] Lung cancer is one of the most common malignant tumors in my country, and its morbidity and mortality rank first in the world, and are increasing year by year. Small-cell lung cancer (SCLC) is the most malignant type of lung cancer, accounting for 15-20% of all lung cancer cases. Specific biological behaviors such as metastasis are highly sensitive to radiotherapy and chemotherapy, and most patients develop rapidly and die within a short period of time. Therefore, it is particularly important to diagnose small cell lung cancer early, formulate a reasonable treatment plan, and evaluate the curative effect. At present, the main methods used for auxiliary diagnosis of lung cancer are chest X-ray, CT, and sputum cytology examination, etc., and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N27/48G01N27/30
Inventor 仲召阳王东卿毅戴楠李梦侠关伟金丰单锦露
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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