High-efficiency antibody fixing method of biological chip

A biochip, antibody immobilization technology, applied in biological testing, material inspection products, instruments, etc., can solve the problems of low spot uniformity, low detection limit, low signal intensity, etc., to solve the problem of weak adsorption and avoid antibody shedding , the effect of increasing the capture capacity

Pending Publication Date: 2019-10-01
TIANJIN RES INST FOR ADVANCED EQUIP TSINGHUA UNIV +1
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  • Description
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  • Application Information

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Problems solved by technology

However, the linkage of monolayer molecules leads to large steric hindrance, high non-specific adsorption, low sample uniformity, poor binding force, low signal intensit

Method used

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  • High-efficiency antibody fixing method of biological chip
  • High-efficiency antibody fixing method of biological chip
  • High-efficiency antibody fixing method of biological chip

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Experimental program
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Effect test

Embodiment 1

[0056] In this embodiment, a high-efficiency antibody immobilization method for biochips specifically includes the following steps:

[0057] (1) Cleaning of biochips: The untreated PS substrate chips were ultrasonically cleaned with deionized water and dried with nitrogen gas.

[0058] (2) Biochip surface activation treatment: the cleaned PS substrate chip was activated for 90 s under the conditions of a plasma oxygen flow rate of 500 mL / min and a generation power of 150 w.

[0059] (3) 3D microstructure construction: the plasma-activated PS substrate chip was placed in a chitosan-acetic acid solution with a mass concentration of 0.5 mg / mL, and treated at 37°C for 60 minutes to obtain a 3D microstructure treatment of biochips.

[0060] (4) Grafting of the coupling layer: the obtained 3D microstructure biochip was placed in a glutaraldehyde solution with a volume fraction of 2%, treated at 37° C. for 30 minutes, washed with deionized water and dried for later use.

[0061] (5...

Embodiment 2

[0070] In this embodiment, a high-efficiency antibody immobilization method for biochips specifically includes the following steps:

[0071] The previous 4 steps of biochip cleaning, biochip surface activation treatment, 3D microstructure construction, coupling layer grafting are the same as those in Example 1.

[0072] (5) Immobilization of antibodies: Using a biochip spotter, spot 2uL of antibodies coupled with fluorescent microspheres to a specific area on the chip surface, and incubate for 2 hours at a temperature of 37°C and a humidity of 80%.

[0073] (6) A fluorescence quantitative analyzer is used to detect the fluorescence intensity value of the spotting area.

[0074] (7) Blocking and elution: Soak the chip in BSA solution with a mass concentration of 1% for 30 minutes, then use 0.5% PBST and deionized water to clean the surface of the chip in turn, blow dry with nitrogen, and detect the spotting area with a fluorescence quantitative analysis instrument the fluoresc...

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Abstract

The invention provides a high-efficiency antibody fixing method of a biological chip. The high-efficiency antibody fixing method of the biological chip comprises the steps of: (1), activating the surface of the biological chip; (2), constructing a 3D micro-structure; (3), performing coupling layer grafting; (4), performing covalent fixing of the antibody; and (5), performing closed elution. The method in the invention solves the problem in the existing polymer chip antibody fixing technology. The method is simple in preparation and low in cost, and is suitable for industrial production. The obtained biological chip has the characteristics of being high in antibody fixing efficiency, good in uniformity, low in background and high in signal to noise ratio, and can be applied to multiple in-vitro detection biological chip carriers.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a high-efficiency antibody fixation method for biochips, which is suitable for fixing antibodies, proteins, nucleic acids, etc., and can be directly applied to the production of in vitro detection biochips. Background technique [0002] As a cutting-edge technology in in vitro rapid diagnosis, biochip technology can realize miniaturized, integrated and automated rapid analysis and detection. Due to its great advantages in terms of sample volume, sample type, reagent convenience, and operator requirements, it is widely used in intensive care units, operations, emergency departments, patients' homes, and chronic disease monitoring. In the application of in vitro detection, biochips can detect most routine clinical indicators. The chip is the basis of detection, and its preparation of biochips is a very important link in product development. [0003] Biochips generally cho...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531G01N33/533G01N33/68G01N21/64
CPCG01N21/6428G01N21/6486G01N33/53G01N33/531G01N33/533G01N33/6854
Inventor 李勇索轶平徐涛
Owner TIANJIN RES INST FOR ADVANCED EQUIP TSINGHUA UNIV
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