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Surface plasma resonance immunosense chip as well as preparation method and application thereof

A surface plasmon and immunosensing technology, which is applied in the field of plasmon resonance immunosensing chips and its preparation, can solve the problems of expensive instruments, fluorescent markers polluting the environment, and restrictions

Inactive Publication Date: 2014-05-14
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the detection of allergens is mainly based on enzyme-linked immunosorbent assay (Enzyme linked immunosorbent assay, ELISA), which is highly sensitive, but the detection process is complicated, the fluorescent markers used pollute the environment, and the instrument is expensive. limit

Method used

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  • Surface plasma resonance immunosense chip as well as preparation method and application thereof
  • Surface plasma resonance immunosense chip as well as preparation method and application thereof
  • Surface plasma resonance immunosense chip as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1 Preparation of Surface Plasmon Resonance Immunosensor Chip

[0077] (1) A circular glass plate with a diameter of 20mm and a thickness of 1mm is used as the substrate, and a gold film with a thickness of 50nm is deposited on the substrate as a solid phase carrier of the surface plasmon resonance biochip;

[0078] (2) Add HS (CH 2 ) 10 COOH (mercaptoundecanoic acid) and 9mM HS(CH 2 ) 6 OH (mercaptohexanoic acid) ethanol solution 4mL, warm bath at 37°C, avoid light, chemically modify the surface of the gold film for 2 hours; then wash with PBS buffer, wash thoroughly for 3 times, each time for 2 minutes, wash off unbound substances; Blow dry with nitrogen at 50°C;

[0079] (3) Fix the above-mentioned solid-phase carrier on the SPR detector;

[0080] (4) Wash the flow cell with PBS buffer 3 times, each time for 2 minutes; add N-hydroxysuccinimide (NHS) and N-ethyl-N'-(dimethylaminopropyl) carbon after the baseline is stable Diimine (EDC) mixed solution w...

Embodiment 2

[0083] Embodiment 2 Preparation of Surface Plasmon Resonance Immunosensor Chip

[0084] (1) A circular glass plate with a diameter of 20mm and a thickness of 1mm is used as a substrate, and a gold film with a thickness of 50nm is deposited on the substrate as a solid phase carrier of a surface plasmon resonance biochip;

[0085] (2) Add HS (CH 2 ) 10 COOH (mercaptoundecanoic acid) and 9mM HS(CH 2 ) 6 OH (mercaptohexanoic acid) ethanol solution 4mL, warm bath at 37°C, avoid light, chemically modify the surface of the gold film for 2 hours; then wash with PBS buffer, wash thoroughly for 3 times, each time for 2 minutes, wash off unbound substances; Blow dry with nitrogen at 50°C;

[0086] (3) Fix the above-mentioned solid-phase carrier on the SPR detector;

[0087] (4) Wash the flow cell with PBS buffer 3 times, each time for 2 minutes; add N-hydroxysuccinimide (NHS) and N-ethyl-N'-(dimethylaminopropyl) carbon after the baseline is stable Diimine (EDC) mixed solution 150 μ...

Embodiment 3

[0090] The preparation method of embodiment 3 surface plasmon resonance immune sensor chip

[0091] (1) A circular glass plate with a diameter of 20 mm and a thickness of 1 mm is used as a substrate, and a gold film with a thickness of 50 nm is deposited on the substrate as a solid phase carrier of a surface plasmon resonance biochip;

[0092] (2) Add HS (CH 2 ) 10 COOH (mercaptoundecanoic acid) and 9mM HS(CH 2 ) 6 OH (mercaptohexanoic acid) ethanol solution 4mL, warm bath at 37°C, avoid light, chemically modify the surface of the gold film for 2 hours; then wash with PBS buffer, wash thoroughly for 3 times, each time for 2 minutes, wash off unbound substances; Blow dry with nitrogen at 50°C;

[0093] (3) Fix the above-mentioned solid-phase carrier on the SPR detector;

[0094] (4) Wash the flow cell with PBS buffer 3 times, each time for 2 minutes; add N-hydroxysuccinimide (NHS) and N-ethyl-N'-(dimethylaminopropyl) carbon after the baseline is stable Diimine (EDC) mixed...

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Abstract

The invention relates to the field of biochips, and particularly relates to a surface plasma resonance immunosense chip as well as a preparation method and application thereof. The surface plasma resonance immunosense chip is prepared by the following steps: fixing a shrimp tropomyosin, shrimp tropomyosin monoclonal antibody ascites or a shrimp tropomyosin monoclonal antibody on a solid phase carrier as a bioprobe; generating surface plasma resonance response by utilizing a gold membrane; modifying sulfydryl on the surface of the gold membrane by utilizing a self-assembly monomolecular layer technique; and fixing the probe on the biochip after the chip is activated. The biochip can be applied to detection of the shrimp tropomyosin monoclonal antibody or the shrimp tropomyosin monoclonal antibody ascites, and detection of the allergen (shrimp tropomyosin), has the advantage of simplicity and convenience in operation, rapidity, high flexibility, no need of being labeled, low cost, simple device, no pollution to the environment and the like, and is hopeful to realize the field real-time detection of a great amount of samples.

Description

technical field [0001] The invention relates to the field of biochips, in particular to a surface plasmon resonance immune sensor chip and its preparation method and application. Background technique [0002] In the 1990s, the development of the Human Genome Project (Human Genome Project, HGP) and related disciplines of molecular biology provided favorable conditions for the emergence and development of biochip and gene chip technologies. Biochip (biochip) is based on the principle of specific interaction between biomolecules, and the biochemical analysis process is fixed on the surface of the chip, so as to realize high-throughput and rapid detection of DNA, RNA, peptides, proteins and other biological components. A protein chip is obtained by immobilizing some non-nucleic acid living substances such as proteins or antigens on microcarriers. The probes on the chip are composed of proteins or the chips act on proteins, which are collectively referred to as protein chips. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N21/552
CPCG01N21/553G01N33/54373G01N33/6803G01N2333/4712
Inventor 李莹钟金钢马骁齐攀
Owner JINAN UNIVERSITY
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