Surface plasma resonance instrument chip based on polyethyleneglycol and method for making same

A technology of surface plasmon and polyethylene glycol, which is applied in the direction of instruments, scientific instruments, material inspection products, etc., can solve the problem of non-specific adsorption of proteins

Inactive Publication Date: 2008-09-10
武晓荣
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The chip of the present invention effectively solve

Method used

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  • Surface plasma resonance instrument chip based on polyethyleneglycol and method for making same
  • Surface plasma resonance instrument chip based on polyethyleneglycol and method for making same
  • Surface plasma resonance instrument chip based on polyethyleneglycol and method for making same

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1, prepare the surface plasmon resonance instrument chip of the 100% carboxyl functionalized polyethylene glycol matrix of 100% initiator density

[0042] According to the method reported in literature 3: Ma, H.; Wells, M.; Bebees, T.; Chilkoti, A.; The gold surface of the glass sheet is cleaned by ozone ultraviolet rays with a UV / Ozone cleaning machine to remove surface impurities; immediately after cleaning, it is immersed in an ethanol solution containing 1 mmol of bromoisobutyric acid-ω-mercaptoundecyl ester, The mercapto group of the initiator reacts with gold, and a chip covered with 100% initiator self-assembled monolayer can be obtained after 24 hours.

[0043] The liquid reservoir, the sensor chamber of the quartz crystal microbalance and the peristaltic pump are connected with pipelines in sequence, the above-mentioned chip is placed in the sensor chamber of the quartz crystal microbalance, and nitrogen gas is supplied for protection for 15 minutes....

Embodiment 2

[0050] Embodiment 2, use the chip that embodiment 1 makes to carry out SPR detection

[0051] In the BIAcore3000 instrument, the chip obtained in Example 1 of this method is used. According to the standard operating procedure, first flow through phosphate buffered saline (PBS buffer, pH=7.4) to obtain the baseline. Because the density of the film obtained by this method is higher, it shows that the RU reaches 30000-40000. The RU of a general dextran chip is only 15,000. Then N-hydroxysuccinimide (NHS, N-hydroxysuccinimide, 0.1mM) and 1-[3-(dimethylamino)-propyl]-3-ethylcarbodiimide hydrochloride (EDC, 1-(3-dimethylaminopropyl )-3-ethylcarbodiimide hydrochloride, 0.1 mM) mixed solution for 30 minutes to activate the carboxyl group, and finally passed bovine serum albumin (BSA). Due to the higher density of COOH obtained by this method, it is shown that more BSA can be connected. Under the same conditions (protein concentration, temperature, time, etc.), the RU obtained by co...

Embodiment 3

[0052] Example 3. Preparation of 60% amino-functionalized polyethylene glycol matrix with 45% initiator density using the method of the present invention

[0053] According to the method reported in literature 3: Ma, H.; Wells, M.; Bebees, T.; Chilkoti, A.; The gold surface of the glass sheet is cleaned with ozone ultraviolet rays by a UV / Ozone cleaning machine to remove surface impurities; immediately after cleaning, it is immersed in bromoisobutyric acid-ω-mercaptoundecyl ester and ω- In the binary mixed ethanol solution of mercaptoundecane (diluent), the molar ratio of the initiator bromoisobutyric acid-ω-mercaptoundecyl ester to the diluent ω-mercaptoundecane is 1:0.4 , the mercapto group reacts with gold, and after 24 hours, a chip covered with a 45% initiator self-assembled monolayer can be obtained (the ratio of the two mercapto compounds in the solution is different from the ratio of the two mercapto compounds on the final surface, which can be Obtained by surface che...

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Abstract

The invention relates to a chip of a surface plasmon resonance instrument which is based on polyethylene glycol and a preparation method. The chip comprises a glass film base and a gold plating film on the glass film base; wherein the gold plating film is provided with an initiator layer which is introduced in the way of self-assembly and a functionalized polyethylene glycol layer which is combined with the initiator layer. The invention uses a quartz crystal microbalance to monitor the growth of the film at the surface of the chip at real time and the chip of the invention effectively solves the problem of non-specific absorption of protein. As the surface ornament of the chip is not limited to traditional carboxylic dextran, the density and types of the functional groups at the chip surface are no longer strictly restricted. Furthermore, the chip of the invention also has good performance in terms of the specificity maintenance and quality control of biological signaling molecule and so on. The preparation method of the chip of the invention has few steps and the low manufacturing cost and is more suitable for industrial production. The chip has more potential of deep development and more superior generality.

Description

technical field [0001] The invention relates to a surface plasmon resonance chip, in particular to a polyethylene glycol-based surface plasmon resonance chip and a preparation method thereof. Background technique [0002] Surface plasmon resonance (Surface Plasma Resonance spectroscopy, hereinafter referred to as SPR) is a sensor for detecting biomolecular interactions. In 1983, Liedberg et al. proposed and verified the idea of ​​SPR as a biosensor for biomedical applications. Based on their series of work, Pharmacia of Sweden launched the first generation of commercialized SPR to the market in 1990. SPR instrument. At present, SPR has been widely used in the detection of biomedicine, clinical diagnosis, new drug development (screening, mechanism research), forensic identification, environmental monitoring, food safety and other fields, and is a standard detection method widely accepted by the scientific and industrial circles. . Compared with the enzyme labeling method, ...

Claims

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Application Information

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IPC IPC(8): G01N21/55G01N33/48G01N21/552
Inventor 马雄明
Owner 武晓荣
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