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Surface plasmon resonance biochip and its preparation method and application

A surface plasmon, biochip technology, applied in the measurement of phase influence characteristics, etc., can solve problems such as tremor, speech disorder, even death, chronic injury, etc.

Inactive Publication Date: 2016-09-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Organophosphorus pesticides will remain in agricultural products such as fruits and vegetables, seriously threatening human health, and have neurotoxic effects on humans and animals, causing neurological disorders, tremors, mental confusion, language disorders, and even death. Poisoning caused by organophosphorus pesticides Incidents occur from time to time, and human beings will accumulate pesticide residues in the body through dietary intake, causing chronic damage to the human body

Method used

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  • Surface plasmon resonance biochip and its preparation method and application
  • Surface plasmon resonance biochip and its preparation method and application
  • Surface plasmon resonance biochip and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 Preparation of surface plasmon resonance biochip

[0099] (1) A round glass sheet with a diameter of 20 mm and a thickness of 1 mm is used as the substrate, and a gold film with a thickness of 50 nm is deposited on the glass substrate as the solid phase carrier of the surface plasmon resonance biochip;

[0100] (2) Inject HS(CH) with a final concentration of 0.1mM into the petri dish 2 ) 10 COOH (mercaptoundecanoic acid) and 0.9mM HS(CH 2 ) 6 4 mL of ethanol solution of OH (mercaptohexanoic acid), warm bath at 37°C and avoid light, chemically modify the surface of the gold film for 2h; then pass in PBS buffer to wash it twice, 2min each time to wash off unbound substances; 50°C Dry with nitrogen;

[0101] (3) Fix the above-mentioned solid phase carrier on the SPR detector;

[0102] (4) Wash the flow cell with PBS buffer twice, 2 minutes each time, add N-hydroxysuccinimide (NHS) and N-ethyl-N'-(dimethylaminopropyl) after the baseline is stable 150μL of carbodiimide (E...

Embodiment 2

[0105] Example 2 Using Surface Plasmon Resonance Biochip to Detect Monoclonal Antibody of Cyclophos by Direct Method

[0106] (1) Establish a standard curve:

[0107] Use PBS buffer to prepare standard solutions of different concentrations of coumaphos monoclonal antibody: the concentrations are 0μg / L, 100μg / L, 500μg / L, 1mg / L, 2mg / L; each standard is based on PBS buffer 100 μL of the solution were respectively injected into the micro-flow cell of the surface plasmon resonance instrument, and immunoreacted with the probe on the surface plasmon resonance biochip prepared in Example 1. The reaction time was set to 6min; the surface of the biochip reaction zone was surfaced Plasma resonance scanning, the scanning range is 1~2°; the change of SPR resonance angle is recorded, and the surface plasmon resonance kinetic curve of the immune reaction of each standard solution is obtained. The concentration of the standard solution is taken as the abscissa and the resonance angle is taken as t...

Embodiment 3

[0116] Example 3 Using surface plasmon resonance biochip to detect coumaphos by suppression method

[0117] (1) Establish a standard curve:

[0118] The coumaphos standard product solution is mixed with the coumaphos monoclonal antibody solution and allowed to stand for 8 minutes. In the mixed solution, the coumaphos monoclonal antibody adopts a fixed working concentration: 10mg / L; the final concentrations of the coumaphos standard are respectively 0μg / L, 50μg / L, 100μg / L, 300μg / L, 500μg / L, 1000μg / L, 3000μg / L and 5000μg / L; based on PBS buffer, the mixed solutions of each standard solution and antibody solution are taken separately 100μL was injected into the micro flow cell of the surface plasmon resonance instrument, and immunoreaction occurred with the probe on the surface plasmon resonance biochip prepared in Example 1. The reaction time was 6min; the surface plasmon resonance biochip reaction area was subjected to surface plasmon Resonance scan, the scan range is 1~2°; the chan...

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Abstract

The invention relates to the field of biochips, and in particular relates to a surface plasma resonance biochip, and a preparation method and applications thereof. According to the biochip, a coumaphos-ovalbumin conjugate is immobilized on a solid-phase vector to obtain a biological probe, surface plasma resonant response is made by a gold film, the surface of the gold film is modified with sulfydryl by a self-assembled monolayer technology, and the probe is fixed on the biochip to obtain the biochip after the biochip is activated. The applications of the biochip comprise the application the surface plasma resonance biochip in detecting coumaphos monoclonal antibodies by adopting a direct method and the application of the surface plasma resonance biochip in detecting coumaphos by adopting an inhibition method. Due to the adoption of the direct detection method, the antibodies can be screened, and researches on immune reaction kinetics can be made; due to the adoption of the competitive inhibition method, the concentration of coumaphos micromolecules can be detected with limit of detection of 25 microgram / liter. The biochip has the characteristics of high speed, real-time performance and field detection capability, and is high in sensitivity, low in cost and free of environmental pollution.

Description

Technical field [0001] The invention relates to the field of biochips, in particular to a surface plasmon resonance biochip and a preparation method and application thereof. Background technique [0002] In the 1990s, the development of the Human Genome Project (HGP) and molecular biology related disciplines provided favorable conditions for the emergence and development of biochips and gene chip technologies. Biochip is based on the principle of specific interactions between biomolecules, which fixes the biochemical analysis process on the surface of the chip, so as to achieve high-throughput 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 a microcarrier. The probes on the chip are composed of proteins or the target of the chip is a protein, which is collectively called a protein chip. [0003] Although biochip technology has made con...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/41
Inventor 李莹钟金钢马骁齐攀
Owner JINAN UNIVERSITY
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