Method of constructing SERS spectral probe for detecting breast cancer marker EGFR phosphorylated tyrosine

A probe detection and phosphorylation technology, used in biological testing, measurement devices, material inspection products, etc., can solve problems such as high price and patient risks, and achieve the effect of improving detection sensitivity, ensuring reliability, and improving linear relationship.

Active Publication Date: 2019-11-05
FUJIAN NORMAL UNIV
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However, it is invasive, risky to t

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  • Method of constructing SERS spectral probe for detecting breast cancer marker EGFR phosphorylated tyrosine
  • Method of constructing SERS spectral probe for detecting breast cancer marker EGFR phosphorylated tyrosine
  • Method of constructing SERS spectral probe for detecting breast cancer marker EGFR phosphorylated tyrosine

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Embodiment Construction

[0031] The present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0032] 1. Reagents Chlorauric acid solution (HAuCl4), sodium citrate (Na3C6H5O7), p-mercaptobenzoic acid (4-MBA), p-mercaptobenzonitrile (4-MB), ascorbic acid, silver nitrate (AgNO3), phosphate buffer solution (PBS, PH=7), phosphorylated serine dipeptide (PS-S), phosphorylated threonine dipeptide (PT-T), single-stranded DNA (S1: AAAATGTCCAAGGCT-SH, S2: TTTTACAGGTTCCGA-SH), Bovine serum albumin (BSA), anti-EGFR phosphorylated tyrosine antibody (Anti-EGFR(Phospho-Tyr1172) antibody), antigen EGFR phosphorylated tyrosine polypeptide (DNPDY(P)QQDFFP), EGFR non-phosphotyrosine acid polypeptide (DNPDYQQDFFP). 2. Preparation of the solution

[0033] The preparation of 1% chloroauric acid solution, the specific steps are as follows: First, dissolve the chloroauric acid solid powder in a beaker with ultrapure water, then pour it into a 100mL volumetric fla...

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Abstract

The invention discloses a method of constructing an SERS spectral probe for detecting a breast cancer marker EGFR phosphorylated tyrosine. A gold plate is adopted as a base plate, a nanoparticle of acore-shell structure is adopted as a substrate for SERS detection, and a sandwich-configuration probe molecule with high selectivity and sensitivity is developed to detect phosphorylation of the breast cancer marker EGFR tyrosine. The spectral intensity of an SERS at a biomolecule Raman quiet region peak position of 2221cm<-1> in the 4-MB Raman spectrum is adopted as an internal standard, intensity normalization treatment is carried out on a spectral signal of the labeled molecule 4-MBA, and the influence of a Raman signal in a biomolecule fingerprint region on quantitative detection can be avoided, so that a linear relationship between the concentration of an EGFR phosphorylated tyrosine solution and the SERS intensity is greatly improved, and the detection sensitivity of the SERS spectral probe is further improved. The 4-MB and the 4-MBA are located in a middle layer of a gold core and a silver shell, it is ensured that the nature, spatial structure, quantity and the like of the SERSspectral probe are not changed, and thus the reliability of data is ensured. The method has a great potential in the terms of ultrasensitive detection of cancer markers.

Description

technical field [0001] The invention belongs to the field of phosphorylation detection of biomedical cancer markers, and in particular relates to a method for constructing a SERS spectral probe to detect phosphorylated tyrosine of breast cancer marker EGFR. Background technique [0002] Breast cancer is the second most common cancer in the world, accounting for 25% of all cancers, and breast cancer is also the most common malignant tumor in women in the world. Women over 45 years old. Epidermal growth factor receptor (EGFR) is one of the specific protein biomarkers of breast cancer. It is a transmembrane glycoprotein located on the cell surface. It is a complex with tyrosine kinase and epidermal growth factor and transforming growth factor (TGFa). Upon binding, activates and induces cell proliferation and differentiation. EGFR is overexpressed in many epithelial tumors (>75.3µg·L -1 ), such as breast cancer, gastric cancer, esophageal cancer, prostate cancer, etc. [...

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/574G01N21/65
CPCG01N33/6872G01N33/57415G01N33/57484G01N33/58G01N21/658
Inventor 冯尚源刘秀杰卢玉栋
Owner FUJIAN NORMAL UNIV
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