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A gold-based extraction material functionalized with different affinity ligands and its application in surface plasmon optical affinity sandwich analysis

A surface plasmon and sandwich analysis technology, which is applied in the fields of functionalized materials, affinity recognition and spectral detection, and biochemical analysis, to achieve high sensitivity

Active Publication Date: 2020-02-07
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the bottleneck problem of sample pretreatment and detection method existing in the analysis of trace components in complex sample systems such as living single cells, living animals, blood, and urine, the purpose of the present invention is to provide a gold compound functionalized with different affinity ligands. Gold-based extraction materials and the surface plasmon optical affinity sandwich analysis method based on the gold-based extraction materials, using the extraction materials and methods to detect proteins, nucleic acids and sugars in living single cells, living animals, serum, urine, etc. Application in the analysis and detection of biomolecules, as well as in other important detection compounds that can be identified by extraction and identification of gold-based extraction materials

Method used

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  • A gold-based extraction material functionalized with different affinity ligands and its application in surface plasmon optical affinity sandwich analysis
  • A gold-based extraction material functionalized with different affinity ligands and its application in surface plasmon optical affinity sandwich analysis
  • A gold-based extraction material functionalized with different affinity ligands and its application in surface plasmon optical affinity sandwich analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Investigation of extraction materials and nano-Raman probe materials

[0046] The material of the extraction material and the material of the nano-Raman probe have a great influence on the Raman signal. Gold and silver are two commonly used noble metals. Therefore, these two metals are used as the extraction material and the Raman probe respectively. A combination of metals with the strongest Mann signal.

[0047] Preparation of extraction materials of different materials: Cut gold wire and silver wire with a diameter of 0.2 mm into small pieces with a length of about 0.5 cm as the grasping substrates of the two materials, and immerse the small pieces of gold wire and silver wire in 150 μL 3- Aminopropyltriethoxysilane (APTES), 150uL of secondary pure water and 3.7mL of absolute ethanol mixed solution, reacted at room temperature for 12 hours, so that the surface of the gold wire and silver wire was modified with amino groups, and then cleaned with ethanol T...

Embodiment 2

[0050] Embodiment 2: the preparation of basic extraction material

[0051] The base extraction material is prepared by acid reduction reaction. Dip different forms of extraction materials (extraction materials include glass rods, glass capillaries, acupuncture needles, metal needles, metal wires, glass flakes, etc.) Reduction at -55°C for 4-5 hours, the surface of the extraction material is modified with a gold thin layer, and the basic extraction material is obtained. The obtained base extraction material was dried at 100° C. under vacuum for 12 hours, and finally stored at room temperature for future use. The overall appearance of the basic extraction material is as follows: figure 2 a, 2c and 2e are shown on the left.

Embodiment 3

[0052] Embodiment 3: Preparation of boric acid functionalized extraction material

[0053] The basic extraction material prepared in Example 2 was immersed in 3mL of a mixed solution consisting of 3-aminopropyltriethoxysilane (APTES), secondary pure water and absolute ethanol in a volume ratio of 5:5:9, at room temperature The reaction was carried out for 12 hours, and after the reaction was completed, the unreacted reagent was removed by washing with absolute ethanol for 3-5 times. An amino-functionalized extraction material is thus obtained.

[0054]Immerse the above-prepared amino-functional extraction material in 40 mL of methanol mixed solution containing 10 mg / mL 4-formylphenylboronic acid and 1% (w / w) sodium cyanoborohydride, and react at room temperature for 24 hours. Wash with secondary pure water and absolute ethanol for 3-5 times to remove unreacted reagents, and then vacuum dry at 40° C. for 12 hours, thereby obtaining boric acid functionalized extraction material...

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Abstract

The invention discloses a gold-based extraction material functionalized by different affinity ligands, and an application thereof in surface Plasmon optical affinity sandwich analysis. The surface of the extraction material is modified with a gold thin layer, and the surface of the gold thin layer is modified with an antibody, a molecularly imprinted polymer, nucleic acid or other affinity identification ligands or materials. A target analyte in a sample to be detected is specifically extracted to the surface of the extraction material by using the gold-based extraction material functionalized by different affinity ligands, the target analyte is labeled by using a silver-based nanometer Raman probe modified with the affinity ligand capable of identifying the target analyte to obtain a sandwich compound formed by the extraction material, the target compound and the Raman probe, and the content of the target analyte is detected through detecting a Raman signal. The extraction material has the advantages of high specificity, effective removal of interference of complex sample matrixes, high detection sensitivity reaching a single-molecular level, simple method and fast analysis speed, and can be used in living single cells, living animal analysis and immunological and biochemical analysis.

Description

technical field [0001] The invention belongs to the fields of biochemical analysis, functionalized materials, affinity recognition and spectral detection, and specifically relates to a gold-based extraction material functionalized with different affinity ligands and its application in surface plasmon optical affinity sandwich analysis . Background technique [0002] The analysis of complex biological samples is often faced in the fields of life science research and clinical diagnosis. The complex composition of biological samples, the abundance of structural analogues, the often extremely low concentration of target analytes, and the severe interference of the sample matrix pose important challenges for life analytical chemistry. There are two key issues in the analysis of complex biological samples: first, the specific enrichment of the target, and second, high-sensitivity detection. Immunoassays, especially enzyme-linked immunoassays, are currently the most common means ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 刘震刘佳尹丹阳涂雪莹皮耶·木合马徳
Owner NANJING UNIV
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