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Biochip for combined detection of multiple tumor markers and preparation and application thereof

A biochip and chip technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of chemical modification on the surface of gold foil chips that have not been found, and achieve the effects of good geometric pictures, control of steric hindrance effects, and stable combination

Pending Publication Date: 2022-01-18
ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, CDI has not been found to be applied to the surface chemical modification of gold foil chips

Method used

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  • Biochip for combined detection of multiple tumor markers and preparation and application thereof
  • Biochip for combined detection of multiple tumor markers and preparation and application thereof
  • Biochip for combined detection of multiple tumor markers and preparation and application thereof

Examples

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preparation example Construction

[0052] In the preparation of the co-modified solid phase carrier of the present invention, the step of cleaning and drying the gold foil chip is also included before the modification, wherein the method of cleaning and drying is as follows: the NH 3 、H 2 o 2 and H 2 O is mixed at a volume ratio of 1:1:5 to form a TL1 cleaning solution. Immerse the gold foil chip in a stainless steel cleaning box filled with TLI cleaning solution, bathe in 82°C water for 6 minutes, rinse it with ultrapure water after taking it out, and then clean it with absolute ethanol , Blow dry with nitrogen.

[0053] The present invention also provides the application of the solid phase carrier in the preparation of in vitro diagnostic products, and the in vitro diagnostic products are biochips or detection kits, or protein arrays.

[0054] In a specific embodiment, the present invention also provides the application of the solid phase carrier in the preparation of biochips. The biochip is selected fro...

Embodiment 1

[0127] Example 1, Molecular Self-Assembled Monolayer Formation and Probe Solidification

[0128] Clean the gold foil chips: NH 3 、H 2 o 2 and H 2 O is mixed at a volume ratio of 1:1:5 to form a TLI cleaning solution, which is placed in a stainless steel cleaning box. Immerse the gold foil chip in a stainless steel cleaning box filled with TL1 cleaning solution, bathe in 82°C water for 6 minutes, take it out and rinse it with ultrapure water 4 times, then soak and clean it with absolute ethanol (3min / time x 2 times), blow dry with nitrogen , put it in a clean and airtight chip box after drying, and store it for later use.

[0129] Molecular self-assembled single-layer chip preparation: soak the above-mentioned dried gold foil chip in a mixed mercaptoacid solution composed of 16-MHDA and 6-MHA (the mixed mercaptoacid solution of 0.01mol / L (16-MHDA / 6-MHA molar ratio= 1:10), at room temperature, taken out after incubation for 24 hours, rinsed with absolute ethanol, and dried ...

Embodiment 2

[0131] Embodiment 2, immunological characterization of the chip

[0132]Take one sheet of the modified molecular self-assembled monolayer chip prepared in Example 1, and from left to right, sequentially add 11-stage gradient dilution working concentrations of 100 μg / mL-0.1 μg / mL [0.01mol / L PBST-BSA buffer solution (pH=7.4)] of human IgG solution (0.01mol / L PBST-BSA buffer was added dropwise on the far side as a blank control; the three lines were three parallels for each group), at room temperature, incubated with shaking for 2 hours, PBST buffer Rinse and dry under nitrogen flow; add 1 μL of Cy3-labeled donkey anti-human IgG antibody (5 μg / mL) dropwise to the human IgG-coated chip, incubate at room temperature for 2 hours in the dark, rinse with PBST buffer, and blow dry with nitrogen .

[0133] The chip scanner LuxscanTM 10K-A (Boao Co., Ltd., China) is used to scan and detect the above chip, wherein the scanning parameters are set to PMT 90 / power650, and the excitation wav...

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Abstract

The invention discloses a novel biochip for detecting multiple tumor markers and preparation and application thereof The tumor markers include but are not limited to AFP, AFP-L3, GPC3, GP73, OPN and the like. According to the biochip, a gold foil chip modified by a mixed sulfhydryl compound and N, N-carbonyl diimidazole is used as a solid-phase carrier, a polyclonal capture antibody probe for resisting multiple markers is fixed on the surface in a dot matrix manner, and a specific monoclonal detection antibody for resisting the markers is added, so that joint detection of the markers in a sample to be detected is realized. The biochip can accurately detect to-be-detected sample markers such as liver cancer markers, has the advantages of high throughput, high sensitivity, high selectivity, high stability, high specificity and the like, is suitable for large-scale population screening and clinical patient auxiliary diagnosis, and is especially suitable for areas with high incidence of liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedical technology detection, and relates to a biochip for joint detection of multiple tumor markers and its preparation and application, in particular to a protein chip for joint detection of liver cancer serum markers and its preparation and application, especially Involvement of a 16-mercaptohexadecanoic acid (16-MHDA), 6-mercaptohexanoic acid (6-MHA) and N,N-carbonyldiimidazole (CDI)-modified hepatocellular carcinoma (HCC) serum in five liver cancers A protein chip for marker detection and its preparation and application. Background technique [0002] As the basic substance of life, protein is the main bearer of life activities and can most directly reflect the functional state of cells and tissues. Therefore, tissue or body fluid protein is often used clinically as a detection index for body organ function changes. For a long time, serum protein markers for liver cancer have been continuously researched a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/553G01N33/543G01N33/574
CPCG01N33/553G01N33/54393G01N33/57438G01N33/57488G01N33/57476
Inventor 杜卫东房永盛
Owner ANHUI MEDICAL UNIV
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