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Bio-sensitive chip as well as preparation method and use thereof

A sensitive chip, biological technology, applied in the biological field, can solve the problems of increasing detection costs and steps, and achieve the effects of low cost, good portability, and improved sensitivity

Inactive Publication Date: 2013-12-25
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this detection method needs to label the target protein to achieve specific detection. The target protein must be labeled before clinical detection, which increases the detection cost and steps. Therefore, it is imperative to develop fast, high-specificity and label-free biosensitive chip technology. Row

Method used

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  • Bio-sensitive chip as well as preparation method and use thereof
  • Bio-sensitive chip as well as preparation method and use thereof
  • Bio-sensitive chip as well as preparation method and use thereof

Examples

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preparation example Construction

[0028] The preparation method of the above-mentioned biosensitive chip and the detection method to the antigen are realized through the following steps:

[0029] a, first detect the ultraviolet maximum absorption wavelength of the silver nanochip;

[0030] b. Soak the silver nanochip in 0.1-5mM 11-mercaptoundecanoic acid absolute ethanol solution at 15-25°C for 6-24 hours, wash with absolute ethanol and ultra-pure water respectively, blow dry with nitrogen and detect ultraviolet light maximum absorption wavelength;

[0031]c. Soak the chip obtained in step b at 15-25°C in an equal-volume mixed aqueous solution of 7.5-150mM EDC.HCl and 1.5-30mM MNHS to activate the surface carboxyl groups for 1-10 hours, wash with ultrapure water, and blow dry with nitrogen Detect the maximum ultraviolet absorption wavelength;

[0032] d. Add monoclonal antibody with a concentration of 1 μg / ml to 30 μg / ml and a volume of 1 μl to 30 μl dropwise on the surface of the chip obtained in step c, re...

Embodiment 1

[0052] Example 1 Preparation of anti-albumin monoclonal antibody biosensitive chip and detection of albumin

[0053] a, first detect the ultraviolet maximum absorption wavelength of the silver nanochip;

[0054] b. Soak the silver nanochip in 1mM 11-mercaptoundecanoic acid ethanol solution for 12 hours at 20°C, wash it with absolute ethanol and ultrapure water respectively, dry it with nitrogen, and detect the maximum ultraviolet absorption wavelength;

[0055] c. Soak the silver nanochip in 75mM EDC.HCl, 15mM NHS equal-volume mixed aqueous solution at 20°C to activate the surface carboxyl groups for 2 hours, wash with ultrapure water, dry with nitrogen, and measure the maximum ultraviolet absorption wavelength;

[0056] d. Add 10 μl of 10 μg / ml anti-albumin monoclonal antibody solution dropwise to the surface of the silver nanochip, overnight in a 4°C refrigerator for 10 hours, wash with PBS with a pH=7.4 and a concentration of 0.01M, blow dry with nitrogen gas, and detect th...

Embodiment 2

[0061] Example 2 Preparation of anti-SCCA monoclonal antibody biosensitive chip and detection of SCCA

[0062] a, first detect the ultraviolet maximum absorption wavelength of the silver nanochip;

[0063] b. Soak the silver nanochip in 1mM 11-mercaptoundecanoic acid ethanol solution for 12 hours at 20°C, wash it with absolute ethanol and ultrapure water respectively, dry it with nitrogen, and detect the maximum ultraviolet absorption wavelength;

[0064] c. Soak the silver nanochip in 75mM EDC.HCl, 15mM NHS equal-volume mixed aqueous solution at 20°C to activate the surface carboxyl groups for 2 hours, wash with ultrapure water, dry with nitrogen, and measure the maximum ultraviolet absorption wavelength;

[0065] d. Add 10 μl of 5 μg / ml anti-SCCA monoclonal antibody solution dropwise on the surface of the silver nanochip, overnight in a 4°C refrigerator for 10 hours, wash with PBS with a pH=7.4 and a concentration of 0.01M, blow dry with nitrogen gas, and detect the maximum ...

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Abstract

The invention belongs to the biological field, and specifically relates to a bio-sensitive chip as well as a preparation method and use thereof. The problem to be solved by the invention is that the invention provides the bio-sensitive chip which comprises a silver nano chip, 11-sulfydryl hendecanoic acid fixed on the silver nano chip and an antibody connected with the 11-sulfydryl hendecanoic acid. The invention further provides a preparation method of the bio-sensitive chip. The preparation method comprises the following steps: fixing the 11-sulfydryl hendecanoic acid on the silver nano chip; activating the 11-sulfydryl hendecanoic acid; and connecting the 11-sulfydryl hendecanoic acid with the antibody. The invention further provides use of the bio-sensitive chip. A target antigen protein is detected through specific reaction of the antigen of the antibody by means of the antibody on the bio-sensitive chip.

Description

technical field [0001] The invention belongs to the biological field, and in particular relates to a biologically sensitive chip, a preparation method and application thereof. Background technique [0002] The successful preparation of biosensitive chips is a key step in realizing the specific detection of target substances by biosensors. Due to the high selectivity of antibodies to target antigens in biological reactions, antibody molecules are often immobilized on the surface of various sensing carriers for immunoassay (Phillips TM, Queen WD, More NS, Thompson AM. Protein A coated glass beads universal support medium for high-performance immunoaffinity chromatography. Journal of Chromatography, 1985, 327: 213-219). Physical adsorption is a traditional technique for immobilizing antibody molecules on the surface of a carrier. However, because antibody molecules are sensitive to changes in solution pH, temperature, ionic strength, and substrate materials, they tend to fall ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 郄明蓉丁一段瑞岐李林袁嘉玲程矗赵倩颖
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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