Preparation method of surface plasmon resonance chip based on hyperbranched zwitterionic polymethylacrylcysteine ​​modification

A polymethacryl-based and methacryl-based technology is applied in the field of ion resonance spectrometer anti-pollution chip preparation, which can solve the problems of surface contamination of the sensor chip, false positives, and reduce the accuracy of quantitative detection, and achieve label-free rapid Detection, high solid load, high stability effect

Active Publication Date: 2019-04-23
丁利 +5
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, when surface plasmon resonance is used for analysis and testing, the surface contamination of the sensor chip is a common problem, and non-specific adsorption from biomolecules or microorganisms in the sample will reduce the accuracy of quantitative detection and produce false positive results

Method used

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  • Preparation method of surface plasmon resonance chip based on hyperbranched zwitterionic polymethylacrylcysteine ​​modification
  • Preparation method of surface plasmon resonance chip based on hyperbranched zwitterionic polymethylacrylcysteine ​​modification
  • Preparation method of surface plasmon resonance chip based on hyperbranched zwitterionic polymethylacrylcysteine ​​modification

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Embodiment 1

[0032] A method for preparing a surface plasmon resonance chip based on hyperbranched zwitterionic polymethacrylcysteine ​​modification, comprising the following steps:

[0033] a) Electron beam evaporation coating technology is used to coat a layer of 2nm thick chromium layer and a layer of 48nm thick gold film on the BK7 glass substrate to obtain a bare gold chip.

[0034] b) Immerse the bare gold chip in a mixed solution of 98wt% concentrated sulfuric acid and 30wt% hydrogen peroxide (volume ratio 7:3), soak for 2 hours at room temperature, then take it out, wash it with deionized water for 3 times, and dry it with nitrogen for later use.

[0035] c) Synthesis of methacrylcysteine ​​monomer: Dissolve 24.98mmol cysteine ​​with 20mL deionized water, add 27.47mmol 2-acrylic acid-2-hydroxy-1,3-propanediester and 29.4 μmol xylylphosphorus, stirred and reacted at 20°C for 2h. The reacted solution was extracted once with ethyl acetate and twice with dichloromethane, and the obtai...

Embodiment 2

[0043] A method for preparing a surface plasmon resonance chip based on hyperbranched zwitterionic polymethacrylcysteine ​​modification, comprising the following steps:

[0044] a) A 3nm thick chromium layer and a 47nm thick gold film were coated on the BK7 glass substrate by electron beam evaporation coating technology, and a bare gold chip was obtained.

[0045] b) Dip the bare gold chip into a mixed solution of 98% concentrated sulfuric acid and 30% hydrogen peroxide (volume ratio 7:3) for 2 hours at room temperature. Then take it out and wash it three times with deionized water, and dry it with nitrogen gas for later use.

[0046] c) Dissolve 24.98mmol of cysteine ​​in 20ml of deionized water, add 27.47mmol of 2-acrylic acid-2-hydroxy-1,3-propanediide and 29.4μmol of xylylphosphine, and stir at 30°C for 1h . The reacted solution was extracted once with ethyl acetate and twice with dichloromethane, and the obtained aqueous monomer solution was lyophilized to obtain a whit...

Embodiment 3

[0054] The hyperbranched zwitterionic polymethylacrylcysteine ​​SPR chip obtained in Example 1 was attached to the prism of the SPR system with asphalt. Phosphate buffer with a pH of 7.4 was used as the mobile phase at a flow rate of 10 μL / min. After the baseline is stable, inject 100 μL of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC, the concentration is 0.4M) and N-hydroxysulfosuccinimide into the quantitative loop A mixed solution of amine (NHS, concentration 0.1M) is used to push the EDC / NHS solution to the surface of the chip through the mobile phase. After 15 minutes, 100 μL of goat immunoglobulin (anti-IgG, 1 mg / mL) was injected, and the anti-IgG solution was pushed to the surface of the chip through the mobile phase. After blocking with ethanolamine, the SPR sensor chip with immobilized anti-IgG was obtained after 30 minutes.

[0055] The hyperbranched zwitterionic polymethacrylcysteine ​​SPR chip obtained above was attached to the prism of the SPR system with...

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Abstract

The invention discloses a hyperbranched amphoteric ion polymethyl propenyl cysteine modification based surface plasmon resonance instrument chip preparation method. A prepared chip comprises a BK7 glass sheet substrate coated with a 2-3nm chromium layer and a 45-50nm gold film layer. A polymethyl propenyl cysteine monomer is obtained through 'thiol-ene' click reaction, an initiator realizes surface modification through polyethyleneimine on the gold film surface, and further monomer polymerization on the gold film surface is initiated through surface initiated atom transfer radical polymerization, so that the hyperbranched amphoteric ion polymethyl propenyl cysteine modification based surface plasmon resonance instrument chip is obtained. The chip prepared according to the method has a remarkable antifouling effect on single protein solution and natural complex sample systems and is high in biomacromolecule loading capacity and high in biomacromolecule compatibility, and high-sensitivity mark-free rapid detection of target molecules in the complex sample systems can be realized.

Description

technical field [0001] The invention relates to the field of preparation of anti-pollution chips for surface plasmon resonance spectrometers, in particular to a preparation method for surface plasmon resonance spectrometer chips based on hyperbranched zwitterionic polymethylacrylcysteine ​​modification. Background technique [0002] The Surface Plasmon Resonance (SPR) instrument is a biosensor technology that appeared in the 1980s. When the incident light is incident from a high-refractive-index medium to a low-refractive-index medium, total reflection will occur. A layer of metal film (gold or silver) is coated at the interface of the medium. The evanescent wave generated by the incident light will cause the free electrons on the metal surface to oscillate collectively, and then form plasma. When the frequency and wave number of the evanescent wave and surface plasmon oscillation When they are equal, the resonance phenomenon (SPR) will occur, which will cause the energy to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/552
CPCG01N21/553
Inventor 丁利苏荣欣叶慧君王利兵齐崴王季诚
Owner 丁利
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