A kind of surface multifunctional coating and its preparation method and application
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A kind of chemical coating and multi-functional technology, applied in the field of analytical chemistry, can solve the problems of anti-non-specific adsorption performance damage, etc., and achieve the effect of strong tolerance, good stability and good versatility
Active Publication Date: 2019-06-25
INST OF CHEM CHINESE ACAD OF SCI
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Literature (Vaisocherova, H.; Sevcu, V.; Adam, P.; Spackova, B.; Hegnerova, K.; Pereira, A.d.l.S.; Rodriguez-Emmenegger, C.; Riedel, T.; Houska, M.; Brynda, E.; Homola, J.Biosensors&bioelectronics 2014,51,150-157.) reported that the polyhydroxy polymer polyhydroxyethyl methacrylate (pHEMA) with good non-specific adsorption properties was treated with N, N'-disuccinimide And carbonate (DSC) / 4-dimethylaminopyridine (DMAP) functionalization for protein immobilization, and then deactivation (blocking) step, its anti-non-specific adsorption performance will be severely damaged
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Embodiment 1
[0061] to immobilize glucose and chicken ovalbumin (e.g. figure 1 shown), the specific preparation steps are as follows:
[0062] 1. Preparation of a functionalized layer with glucose and chicken ovalbumin on the surface of gold membrane
[0063] 1. In situ modification initiator of gold film surface
[0064] Soak the gold film in 10mmol / L, 0.75mL ethanol solution of mercaptoundecanol (MUOH), react at 4°C for 12h, wash with ethanol three times, and blow dry with nitrogen. The gold film was placed in a suitable reactor, 1 mL of tetrahydrofuran (13.7 mmol) and 84 μL of pyridine (1.0 mmol) were added, and 124 μL of 2-bromoisobutyryl bromide (BIBB,) (1.0 mmol) was added dropwise. React at room temperature at 25°C for 30 minutes, wash the gold film with THF three times, and dry it with nitrogen.
[0065] 2. In-situ polymerization of polyhydroxyethyl methacrylate (pHEMA, molecular weight about 30,000) on the surface of the gold film
[0066] The in situ polymerization of pHEMA n...
Embodiment 2
[0076] Lysine is immobilized on the glass surface, and the preparation steps of the specific functionalized layer are as follows:
[0077] 1. Preparation of functionalized layer with lysine on glass surface
[0078] 1. In situ modification initiator of glass surface
[0079] After plasma cleaning (70W, 3min), the glass substrate was immediately immersed in 0.75mL, 3% (mass fraction) 3-amino-trimethoxysilane (ATPMS) toluene solution to react for 30min, and then ultrasonically cleaned with toluene and acetone for 3 times respectively. , blow dry with nitrogen. The gold film was placed in a suitable reactor, (13.7 mmol) tetrahydrofuran and 1.0 mmol) pyridine were added, and 1.0 mmol 2-bromoisobutyryl bromide (BIBB) was added dropwise. After reacting for 30 minutes, the glass substrate was washed three times with tetrahydrofuran, and dried with nitrogen.
[0080] 2. In situ polymerization of pHEMA on the glass surface
[0081] The in situ polymerization of pHEMA needs to be ca...
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Abstract
The invention discloses a surface multi-functionalized coating and a preparation method and application thereof. The surface multi-functionalized coating is formed by connecting a substrate, a polyhydroxy compound, bridging molecules, probe molecules and deactivated molecules, the polyhydroxy compound is modified on the substrate and connected with the probe molecules through the bridging molecules, and residual reaction loci of the bridging molecules after being connected with the polyhydroxy compound and the probe molecules are connected with the deactivated molecules. The preparation method includes following steps: modifying the polyhydroxy compound on the substrate; being in nucleophilic substitution with the bridging molecules; enabling the bridging molecules after reaction to be in nucleophilic substitution with the probe molecules; adding the deactivated molecules for reaction to obtain the surface multi-functionalized coating. The preparation method has the advantages of simple operation, easiness and flexibility in control and low modifying cost, and the surface multi-functionalized coating obtained by the method has the advantages of remarkable effect, high stability and little interference to biomolecule recognition.
Description
technical field [0001] The invention relates to a surface multifunctional coating and its preparation method and application, belonging to the field of analytical chemistry. Background technique [0002] The surface of the sensor chip usually needs to be functionalized for the immobilization of probe molecules. However, for sensor chips such as the gold film of SPRi, the sites of target molecules that can be immobilized per unit area are fixed, limited and related to the size of the immobilized molecules. Taking the immobilization of small molecule mannose as an example, if the mannose molecule is reduced to a cube, the simulated value of its side length is about Up to about 3 molecules can be confluent on each square nanometer of surface. As a result, the sensitivity of SPR or SPRi assays is severely lacking, often with no detectable signal. In order to improve detection sensitivity, it is necessary to increase the amount of probe immobilized per unit area. This requir...
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