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Method for detecting low-abundance protein in blood plasma through surface-enhanced Raman spectroscopy (SERS)

A technology of surface-enhanced Raman and low-abundance proteins, which is applied in the field of low-abundance proteins, can solve the problems of inability to identify and distinguish proteins of equal mass, high cost of mass spectrometers, and difficulty in widespread promotion, so as to make up for the identification of low-abundance proteins by mass spectrometry The lack of protein, the enrichment process is simple and easy, and the effect of reducing costs

Active Publication Date: 2014-01-29
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods all have the problems of large sample volume, expensive consumables, complicated steps, and long time-consuming.
In addition, mass spectrometry is easily interfered by eluent and other factors, and it is impossible to identify and distinguish proteins of equal mass. Moreover, mass spectrometers are expensive and difficult to be widely used.

Method used

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  • Method for detecting low-abundance protein in blood plasma through surface-enhanced Raman spectroscopy (SERS)
  • Method for detecting low-abundance protein in blood plasma through surface-enhanced Raman spectroscopy (SERS)
  • Method for detecting low-abundance protein in blood plasma through surface-enhanced Raman spectroscopy (SERS)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. SERS spectrum detection of low-abundance proteins in plasma of liver cancer patients based on silver nanoparticles enhanced substrate

[0044] (1) Removal of high-abundance proteins in the plasma of patients with liver cancer

[0045] Take 2 g of ion exchange resin in a collection tube covered with a spin column, add 100 μl of eluent, mix well, centrifuge at 3000 rpm for 5 min, pour out the liquid in the collection tube, and repeat three times. Take 10 μl of liver cancer patient plasma standard sample and dilute it in 100 μl of eluent, mix it evenly, add it to the spin column containing ion exchange resin, use a pipette to repeatedly suck and mix well, place it on a shaker at 200rpm for 15min, and shake it at 2000rpm The plasma low-abundance protein solution of patients with liver cancer was collected by centrifugation.

[0046] (2) Enrichment of low-abundance proteins in plasma of patients with liver cancer

[0047] Take 0.2 g of cellulose acetate membrane and inc...

Embodiment 2

[0055] SERS Spectral Detection of Low Abundance Proteins in Plasma of Nasopharyngeal Carcinoma Patients Based on Gold Nanoparticles Enhanced Substrate

[0056] (1) Removal of high-abundance proteins in the plasma of patients with nasopharyngeal carcinoma

[0057] Take 25g of ion exchange resin in a collection tube covered with a spin column, add 1000μl of eluent and mix well with the hydrophobic polymer, centrifuge at 6000rpm for 1min, pour out the liquid in the collection tube, and repeat three times. Take 100 μl of nasopharyngeal carcinoma patient plasma standard sample and dilute it in 1000 μl of eluent, mix it evenly, add it to the spin column containing ion exchange resin, use a pipette to repeatedly pump and mix well, place it on a shaker at 350rpm for 10min , centrifuge at 5000rpm to collect the plasma low-abundance protein solution of patients with nasopharyngeal carcinoma.

[0058] (2) Enrichment of low-abundance proteins in plasma of patients with nasopharyngeal car...

Embodiment 3

[0067] 1. SERS spectrum detection of low-abundance proteins in plasma of gastric cancer patients based on silver nanoparticles enhanced substrate

[0068] (1) Removal of high-abundance proteins in the plasma of patients with gastric cancer

[0069] Take 2 g of silica gel in a collection tube covered with a spin column, add 150 μl of eluent, mix well, centrifuge at 3000 rpm for 5 min, pour out the liquid in the collection tube, and repeat three times. Take 10 μl of gastric cancer patient plasma standard sample and dilute it in 100 μl of eluent, mix it evenly, add it to the spin column containing silica gel, use a pipette to repeatedly suck and mix well, place it on a shaker at 200rpm for 15min, and collect it by centrifugation at 2000rpm Plasma low-abundance protein solution in patients with liver cancer.

[0070] (2) Enrichment of low-abundance proteins in plasma of patients with gastric cancer

[0071] Take 2g of nitrocellulose membrane and incubate with the plasma low-abun...

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Abstract

The invention relates to a method for detecting a low-abundance protein in blood plasma through surface-enhanced Raman spectroscopy (SERS). The method comprises the following steps: adding a standard blood plasma sample into a centrifugal column of a hydrophobic polymer, performing vibration in a shaking table, and performing centrifugation to obtain a low-abundance protein solution; mixing the low-abundance protein solution with a blotting immobilized matrix and the standard blood plasma sample, and performing incubation to obtain an enrichment blood plasma low-abundance protein blotting immobilized matrix; adding glacial acetic acid and SERS active metal nano-particles; performing incubation to obtain a low-abundance protein SERS enhanced matrix mixed solution; performing SERS detection on the mixed solution; through the operation, acquiring blood plasma low-abundance protein SERS data of a cancer patient with an unknown cancer type; and determining the unknown cancer type by comparing the data with Raman spectral databases of healthy people and known cancer types. The method has the advantages of short detection time, low required power and good economic and social benefits; the immobilized matrix is low in cost; an enrichment process is simple and easy to operate.

Description

technical field [0001] The invention relates to the field of protein analysis and detection, in particular to a method for detecting low-abundance proteins in plasma by using surface-enhanced Raman spectroscopy (SERS). Background technique [0002] Raman spectroscopy is a kind of molecular vibration spectrum. It is an analytical method that analyzes the scattering spectrum different from the incident light frequency to obtain molecular vibration and rotation information, and is applied to the study of molecular structure. The Raman spectroscopy analysis process is simple, fast, and highly sensitive. It is a non-destructive testing method. However, the Raman spectroscopy signal is weak and easily interfered by autofluorescence. Therefore, there are certain limitations in direct detection using Raman spectroscopy. Surface-enhanced Raman spectroscopy (SERS) technology is a commonly used Raman signal enhancement method. When molecules are adsorbed on the surface of some rough me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
Inventor 林居强陈荣黄祖芳王静陈冠楠冯尚源李永增谢树森
Owner FUJIAN NORMAL UNIV
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