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Antibody chip for detecting colorectal cancer related factors

An antibody chip, colorectal cancer technology, applied in measuring devices, biological testing, material inspection products, etc., can solve the problem of single detection index, achieve simple operation, fast detection, and save detection time

Inactive Publication Date: 2018-08-17
WAYEN BIOTECHNOLGIES SHANGHAI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The antibody chip in this study can detect multiple indicators at the same time, overcomes the shortcomings of the existing technology such as cumbersome operation and single detection indicator, and has the advantages of convenient operation, accuracy, high throughput, less sample consumption, and can be popularized and scaled in ordinary laboratories Advantages of production and testing

Method used

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  • Antibody chip for detecting colorectal cancer related factors
  • Antibody chip for detecting colorectal cancer related factors
  • Antibody chip for detecting colorectal cancer related factors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The preparation of embodiment 1 antibody chip

[0049] (1) Antibody screening: Western blot experiments were used to identify the specificity and titer of the antibodies corresponding to the three preferred proteins, and to screen high-quality antibodies.

[0050] (2) Purified antibody: use Replace the antibody solvent with PBS in Ultra 10K ultrafiltration tubes, adjust the antibody concentration to 2mg / ml, aliquot 5ul, and store at -20°C.

[0051] (3) Prepare spotting plate: add 18ul spotting buffer (PBS) to 2ul antibody, mix well, and add to spotting plate; add 18ul positive control (biotin-labeled secondary antibody) and 18ul negative control (PBS) to spotting plate .

[0052](4) Spotting: According to the operating requirements of the chip spotting instrument, do a good job of preparation before spotting, including lotion, system fluid, humidity, temperature, spotting needle, substrate and spotting plate, etc. After sample application, the chip was stored in a h...

Embodiment 2

[0054] Example 2 Application of antibody chip to detect multiple indicators in samples

[0055] 1. Sample Preparation

[0056] Let the venous blood stand for 20 minutes, centrifuge at room temperature at 3000r / min, separate the serum, and use it immediately; or divide it and store it in a -80°C refrigerator. Repeated freezing and thawing should be avoided. Thawed samples should be centrifuged again before testing.

[0057] 2. Biotin labeling of samples

[0058] (1) Sample centrifugation: centrifuge the sample at 4°C, 13,200 rpm for 10 minutes, and transfer the supernatant to a new centrifuge tube;

[0059] (2) Sample labeling: Take 3ul of each sample, put it into a centrifuge tube, add 2ul Biotin / DMF, and use the labeling buffer to make up the final volume of the reaction to 70ul. Mix well, shake and react at room temperature for 2h. Add 30ul of stop solution, shake at room temperature for 30min.

[0060] 3. Preparation of Standards

[0061] (1) Add 500ul sample diluent ...

Embodiment 3

[0080] Example 3 Sample Detection Specificity and Sensitivity Analysis

[0081] Using 95 cases of serum samples, the specificity and sensitivity of the kit of the present invention were tested. The detection cutoff value of CEA was 2479pg / ml, that of IGFBP-2 was 7785pg / ml, and that of IL-8 was 445pg / ml. Receiver operator characteristic curve (ROC) analysis was performed using Medcalc software to obtain data such as sensitivity, specificity and area under the curve (AUC). The sensitivity, specificity and AUC of each detection index to the detection of colorectal cancer serum samples are shown in Table 3. image 3 with Figure 4 .

[0082] Table 3 The sensitivity, specificity and AUC of each detection index to the detection results of colorectal cancer serum samples

[0083] Detection Indicator

[0084] It can be seen from the above table that the performance of the joint detection of the three indicators is better than that of each indicator alone.

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Abstract

The invention relates to the field of biological science, and provides a chip technology for detecting colorectal cancer related antibodies and an antibody chip detection kit. An antibody chip comprises a substrate-based solid phase antibody chip and a microbead or microgel-based liquid suspension chip. A protein chip technology is used to fix specific antibodies related to colorectal cancer, andmultiple antigenic proteins related to the colorectal cancer in a sample can be simultaneously detected. The kit includes the antibody chip, a standard product, a sample diluent, a blocking solution,a labeling solution, a detecting solution, and a washing solution. Through the comprehensive calculation of several detection indicators, an auxiliary means for the detection and analysis of the colorectal cancer related antibodies is provided. Compared with a kit for detecting a single index, the antibody chip detection kit for detecting the several detection indicators has the characteristics ofhigh specificity and high sensitivity, and the antibody chip also has the performance of small sample using amount. The invention also discloses detection methods using the antibody chip, and the detection methods comprise a fluorescence detection method based on sample labeling, a double-anti-sandwich method, and a detection method using the liquid suspension chip.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to an antibody chip for detecting proteins related to colorectal cancer and an antibody chip detection kit. Background technique [0002] Colorectal cancer is one of the most common diseases today, and molecular testing is an approach that has developed rapidly in recent years. At present, a large number of studies have reported a variety of biomarkers related to colorectal cancer, mainly including proteins and nucleic acids, and there are also methods for detecting lipids (patent 201210003024.3). Direct serum detection has the advantages of simple and fast operation, non-invasiveness, and low cost. Moreover, a large number of studies have found that various factors may be related to colorectal cancer (Table 1). [0003] Table 1 Factors associated with colorectal cancer [0004] protein abbreviation Chinese translation BDNF brain derived nerve ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57446G01N33/68
Inventor 张庆华李文娜张春秀宋凯
Owner WAYEN BIOTECHNOLGIES SHANGHAI INC
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