Preparation method of electrochemistry immunosensor for unmarked carcinoembryonic antigen detection

An immunosensor and carcinoembryonic antigen technology, which is applied in the field of preparation of high-throughput electrochemical impedance immunosensors, can solve the problems of easy hydrolysis and high requirements on reaction conditions, and achieve the effects of strong specificity, high detection sensitivity and low detection limit.

Inactive Publication Date: 2013-09-11
ZHEJIANG UNIV
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Problems solved by technology

However, EDC/NHS has high requirements on the reaction conditions, such as the acidity and alkalinity of the

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  • Preparation method of electrochemistry immunosensor for unmarked carcinoembryonic antigen detection
  • Preparation method of electrochemistry immunosensor for unmarked carcinoembryonic antigen detection
  • Preparation method of electrochemistry immunosensor for unmarked carcinoembryonic antigen detection

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[0016] The present invention uses the stable Au-S bond formed by one sulfur bond of gold and hexamethylene dithiol (HDT), and the other sulfur bond of HDT is in turn with N-hydroxysuccinimide-polyethylene glycol-maleyl Imine crosslinker (NHS-(PEG) n -Maleimide Crosslinkers, preferably SM(PEG) 6 ) The maleimide (maleimide) group is covalently bonded, followed by SM (PEG) n The N-hydroxysuccinimide ester (NHS-ester) group forms a stable amide bond with the amino group on the antibody. Compared with the traditional EDC / NHS activation reaction of thiol terminal carboxyl group, SM(PEG) n The reaction time with HDT is fast, and SM(PEG) n It is not easy to be hydrolyzed, the reaction is carried out in a neutral environment at room temperature, and the success rate of the reaction is high, so the efficiency of immobilizing the antibody will also increase. The lower limit of the sensitivity of the electrochemical immunosensor of the present invention is 0.1 fg / ml. By fixing different ty...

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Abstract

The invention discloses a preparation method of an electrochemistry immunosensor for unmarked carcinoembryonic antigen detection. The electrochemistry immunosensor is a planar gold electrode array processed by using a standard MEMS process. During preparation, hexanedithiol and an N-hydroxysuccinimide-polyethylene glycol-maleimide cross-linking agent are sequentially adopted to treat a gold electrode, carcinoembryonic antigen is assembled to capture antibody, and then a bovine serum albumin PBS buffer solution with a mass concentration of 1% is adopted to block to obtain the electrochemistry immunosensor. According to the present invention, the carcinoembryonic antigen capture antibody is immobilized on the gold electrode so as to be used for carcinoembryonic antigen detection; and experiment results show that antibody can be stably and simply immobilized on the surface of the gold electrode with the electrochemistry immunosensor constructed by using the method, wherein the electrochemistry immunosensor has characteristics of high detection sensitivity, low detection limitation and strong specificity.

Description

technical field [0001] The invention relates to a preparation technology of a sensing device, in particular to a preparation method of a high-throughput electrochemical impedance immunosensor for unlabeled carcinoembryonic antigen detection. Background technique [0002] Immunological detection is mainly a means of detection using the specific reaction of antigens and antibodies. Because it can use isotopes, enzymes and chemiluminescent substances to amplify and display detection signals, it is often used for trace substances such as proteins and hormones. detection. The electrochemical immunoassay method has attracted people's attention due to its advantages of high sensitivity, no label, and short analysis time. The key point of the electrochemical immunosensor is the immobilization of the capture antibody corresponding to the detection substance on the surface of the electrode. A common approach is to immobilize antibodies on electrodes using self-assembled monolayers. ...

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Application Information

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IPC IPC(8): G01N33/53
Inventor 王平周洁杜丽萍邹玲胡宁
Owner ZHEJIANG UNIV
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