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Purpose of IGFBP3 in preparation of kit for quantitative detection of liver cancer markers

A quantitative detection and marker technology, used in biological testing, measuring devices, material testing products, etc., can solve the problems of low solubility of recombinant proteins, unfavorable kit preparation, unfavorable production and development, etc., and achieve high sensitivity and easy coating. , the effect of strong binding ability

Active Publication Date: 2019-05-10
RAYBIOTECH INC GUANGZHOU +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in order to apply IGFBP3 to the diagnosis and monitoring of tumors, the IGFBP3 recombinant protein sold abroad is expensive, which is not conducive to further production and development
However, the IGFBP3 recombinant protein developed by domestic manufacturers has low solubility, which is not conducive to the preparation of kits

Method used

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  • Purpose of IGFBP3 in preparation of kit for quantitative detection of liver cancer markers
  • Purpose of IGFBP3 in preparation of kit for quantitative detection of liver cancer markers
  • Purpose of IGFBP3 in preparation of kit for quantitative detection of liver cancer markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Screening of Liver Cancer Markers

[0029] The experimental volunteers in this example include cancer patients, other cancer patients and healthy people, and the screening process of liver cancer markers refers to figure 1 shown. Based on the ELISA method, the serum samples provided by the experimental volunteers were fully exposed to the antibody chip coated with antibodies against 274 serum markers, and the artificial neural network analysis was performed on the test results of the antibody chip. Set 78 liver cancer samples, 40 other cancer samples and 80 healthy samples to normalize the data provided by the antibody chip. The standard product curve of the target factor is obtained by using the relevant standard product, and the protein concentration is determined according to the standard product curve. It was found that the content of 21 factors in liver cancer samples was significantly different from that in other cancer samples and healthy samples. Ac...

Embodiment 2

[0035] Example 2 Reverse protein chip verification of the detection effect of 6 factors

[0036] The operation steps of the reverse protein chip are as follows:

[0037] Sample processing: Serum samples provided by the experimenter were treated with appropriate buffer and serial dilution.

[0038] Preparation of standard products: prepare 6 factor recombinant proteins of β2-microglobulin, IGFBP3 protein, GP73 protein, GDF15 protein, OPN protein and AFP protein to a concentration of 100g / ml (stock solution), according to the initial standard curve of each factor The concentration was diluted 5 times according to different 1 / 3 times, and a blank control was set.

[0039] Membrane preparation: Spot the diluted serum samples, standard, positive control and blank control onto the membrane. 800cw-Streptavidin was used as a positive control, and PBS buffer containing 1% BSA was used as a negative control. After spotting, the membranes were naturally dried and stored at -80°C. Mem...

Embodiment 3

[0050] The preparation of the stable IGFBP3 standard substance of embodiment 3

[0051] The IGFBP3 protein is a protein consisting of 291 amino acids, and its full-length amino acid sequence is shown in SEQ ID NO:1. The standard curve of IGFBP3 concentration can be determined by using the IGFBP3 standard calibration. The source of the IGFBP3 standard can be the blood sample of the tumor patient or the gene recombinant expression. The IGFBP3 standard can be selected from the purity greater than 95%: IGFBP3 protein, recombinant IGFBP3 Protein, the full-length or fragment comprising the amino acid sequence of SEQ ID NO: 1 or its complex coupled with other groups and other derivatives. The choice of IGFBP3 standard with high solubility is beneficial to the acquisition of experimental data. The standard curve of IGFBP3 concentration was obtained by using the known concentration of IGFBP3 standard, the corresponding curve of the concentration determined by ELISA method and the meas...

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Abstract

The invention relates to a purpose of IGF BP3 in preparation of a kit for quantitative detection of liver cancer markers. The fact that the content change of the IGF BP3 protein and the state of livercancer are obviously related is discovered, and the kit capable of accurately detecting the content of the IGF BP3 protein is provided so that the diagnosis of the existence, staging and transfer ofliver cancer are assisted.

Description

technical field [0001] The invention belongs to the field of tumor diagnosis, and relates to the use of IGFBP3 in preparing a kit for quantitatively detecting liver cancer markers. Background technique [0002] People talk about cancer discoloration. Since the 21st century, the threat of malignant tumors to human health has become increasingly serious. Its fatality rate is second only to cardiovascular and cerebrovascular diseases, ranking third. The main reason for the high mortality rate of cancer patients is the inability to diagnose early. Early diagnosis and early treatment are the most effective ways to prevent and treat cancer and reduce mortality. [0003] With the development of tumor cells, certain proteins in the body of tumor patients will change, or new abnormal proteins associated with tumors will be produced. These chemical substances that can reflect the presence of tumors are collectively called tumor markers. Tumor markers are antigens and other biologi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/574
Inventor 黄若磐匡治州黄伟毛应清
Owner RAYBIOTECH INC GUANGZHOU
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