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Protein chip for detecting esophageal squamous carcinoma marker and kit box of protein chip

A protein chip and esophageal squamous cell carcinoma technology, which is applied in the field of protein chips to achieve the effects of high sensitivity, reduced cost and improved detection efficiency

Active Publication Date: 2013-06-12
JIANGSU YUANHUA BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In short, the existing technology lacks a protein chip capable of high-throughput screening and detection of esophageal squamous cell carcinoma, the price is acceptable to people, and can be popularized and promoted in population examinations

Method used

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  • Protein chip for detecting esophageal squamous carcinoma marker and kit box of protein chip
  • Protein chip for detecting esophageal squamous carcinoma marker and kit box of protein chip
  • Protein chip for detecting esophageal squamous carcinoma marker and kit box of protein chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of a protein chip for detecting esophageal squamous cell carcinoma

[0031] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the dot array and the spotting position; (3) The spotting needle sucks out the antigen and spots it by spraying, and the electronic voltage spray technology simultaneously Spray onto a glass substrate, (4) assemble a nitrocellulose membrane, and seal, dry, pack, and store.

[0032] The preparation method of the above-mentioned protein chip for detecting various serum antibody markers of esophageal squamous cell carcinoma specifically includes the following steps:

[0033] 1. The source of antigen and antibody for esophageal squamous cell carcinoma marker detection

[0034] (1) Determination of the detection index antigen:

[0035] All antigens were purchased from OriGene, Abnova, and MyBioSource.

[0036] (2) Determination of the detection index antibody:

[0037]...

Embodiment 2

[0043] Kit preparation

[0044] A kit for detecting serum and / or plasma markers for esophageal squamous cell carcinoma, such as Figure 4 As shown, it consists of sample diluent 4, concentrated washing solution 6, bovine serum albumin solution 9, biotin-labeled human IgG antibody 11, anti-biotin-labeled gold cross-linking reagent 12, and detection solution AB (7, 8). All liquids are bottled, stored at 4°C, and used together for inspection.

[0045] Sample Diluent 4 is a protein stabilizer; it can be formulated from 3% BSA and 0.05% detergent (pH=7.3). Purchased from Gentel Company (Gentel Dilution Buffer TM, product number: No.B255). Supplied in 1 bottle, 500ml. Used to dilute serum samples (volume ratio 1:500 or 1000).

[0046] Concentrated Wash Solution 6 is a neutral buffer solution (10×Gentel Wash Buffer TM, product number: No.2-1016); it can be prepared from detergent (pH=6.8). Provide 250mL, one bottle. Dilute with 25mL to 225mL ultrapure water and mix well.

[00...

Embodiment 3

[0052] Application and detection of protein chips and kits

[0053] The application of the protein chip for detecting esophageal squamous cell carcinoma marker antigen according to the present invention mainly adopts the method of antigen-antibody-secondary antibody-nano gold binding-silver deposition method to detect the esophageal squamous cell carcinoma marker antigen in human blood, the method is Add human serum to the surface of the chip, and the antibodies in the sample are bound to the corresponding antigens immobilized on the chip respectively. After drying, add the secondary antibody to react, wash away the unbound secondary antibody, add gold cross-linking reagent, and combine with the secondary antibody , wash off the unbound nano-gold, add silver nitrate, the nano-gold promotes the deposition of silver on the gold surface, scan and collect the signal, the gray level of the sample point is proportional to the concentration of the antibody in the serum.

[0054]In th...

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Abstract

The invention discloses a high-throughput multi-index protein chip for screening esophageal squamous carcinoma specific protein markers, and discloses a kit supporting the use. 55 types of esophageal squamous carcinoma differential proteins in the serum or blood plasma are selected and used as the esophageal squamous carcinoma protein markers and protein contrast prepare the protein chip; and the protein chip comprises a substrate, protein detection indicators and a contrast detection indicator coating, wherein the protein detection indicators are distributed as arrays on the substrate. Detection liquid of a supplementary experiment reagent is filled in the kit. By adopting the protein chip, the protein profiles of three types of people, namely, normal people, people subjected to esophageal squamous cell carcinoma precancerous lesions, and people subjected to the esophageal squamous carcinoma, can be determined; and means are provided for screening the esophageal squamous carcinoma in the early stage, diagnosing in mid stage and late stage, as well as monitoring the state of illness.

Description

[0001] technical field [0002] The invention belongs to the technical field of biochips, in particular to a protein chip. The protein chip and its kit were used to screen for specific protein markers in esophageal squamous cell carcinoma. [0003] Background technique [0004] Esophageal cancer is one of the eighth most common malignant tumors in the world, and is considered to be the second most difficult to treat digestive tract malignant tumors after pancreatic cancer. China is also a country with high morbidity and mortality of esophageal cancer in the world, accounting for more than 50% of the world. There are about 250,000 newly diagnosed cases of esophageal cancer every year. The incidence rate is second only to gastric cancer, lung cancer and liver cancer. Four, a serious threat to human life and health. Esophageal cancer is divided into esophageal squamous cell carcinoma and adenocarcinoma, and the proportion of esophageal squamous cell carcinoma in China is as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 黄桂玲赵伟孙桂菊成于飞钱振宇刘璇
Owner JIANGSU YUANHUA BIO TECH
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