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Biological chip adopting soybean peroxidase (SBP) for marking, and preparation method thereof

A catalase and biochip technology, applied in the biological field, can solve problems such as unstable chemiluminescence signal, and achieve the effects of improved detection sensitivity, high thermal stability and reduced demand

Inactive Publication Date: 2015-01-28
SOUTHEAST UNIV
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Problems solved by technology

[0005] Provides a soybean catalase-labeled biochip, using soybean peroxidase as the labeling enzyme, which solves the problem in the prior art that the chemiluminescent signal generated by the luminol oxidation reaction caused by HRP catalysis is unstable

Method used

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  • Biological chip adopting soybean peroxidase (SBP) for marking, and preparation method thereof

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Embodiment

[0032] Adopt the biochip provided by the present invention to detect four serological markers in the Down's syndrome screening in the second trimester, and its implementation process is as follows: figure 1 shown.

[0033] The PBS involved in this example is 10 mM pH 7.4, Tween20 / TBS ​​solution (Tween20 / TBS, TTBS), 0.3% Tween20 containing 100 mM pH 7.5 Tris-HCL preparation; the soybean catalase used was purchased from Bio -Research Products Company; biotin-labeled antibody was purchased from Shanghai Ruiqi Biotechnology Co., Ltd.; streptavidin was purchased from Sangon Biotech.

[0034] The first step, preparation of enzyme-labeled streptavidin

[0035] Using the improved sodium periodate method: 1. Dissolve 1 mg of soybean catalase in 100 μL of PBS, and add newly prepared 12 mg / mL NaIO 4 100 μL of the solution, mix well, and react at room temperature for 15 minutes in the dark; 2. Add 100 μL of 1% ethylene glycol to the solution obtained in 1 and react at room temperature f...

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Abstract

The invention discloses a biological chip adopting soybean peroxidase (SBP) for marking, and a preparation method thereof. The chip includes a nitrocellulose membrane coated by captured monoclonal antibodies, a biotin-marked polyclonal antibody, SBP-marked streptavidin, and an enhanced chemiluminescent solution where SBP acts, wherein the captured monoclonal antibodies and the biotin-marked polyclonal antibody can all achieve specific binding with an antigen to be detected. The invention has the advantages that the SBP replaces horseradish peroxidase in the prior art to be a marker enzyme for the biological chip preparation, so as to maintain intense chemiluminiscence for a longer time, and achieve much higher detection sensitivity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a biochip labeled with soybean catalase and a preparation method thereof. Background technique [0002] Biochip is a rapidly developing high-tech in the field of life sciences. It mainly refers to a micro-biochemical analysis system constructed on the surface of a solid chip through micro-processing technology and microelectronics technology, in order to realize the analysis of cells, proteins, DNA and other organisms. Accurate, fast, and high-throughput detection of components, it has the advantages of small volume, low consumption of reagents and energy, fast sample analysis speed, and high throughput. The advantages of high-throughput biochips can be used to realize the simultaneous screening of multiple serological markers, and the application of biochip technology to the field of prenatal screening has broken through the clinical screening of Down's syndrome that can only be mea...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/577
Inventor 赵芳刘松琴赵岳五米利卢菊生俞佳超
Owner SOUTHEAST UNIV
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