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
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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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