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Novel application of osteopontin polypeptide

A technology of osteopontin and standard products, applied in the new application field of osteopontin polypeptide

Active Publication Date: 2020-08-04
RAYBIOTECH INC GUANGZHOU +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there are currently no products using OPN for liver cancer detection on the market

Method used

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  • Novel application of osteopontin polypeptide
  • Novel application of osteopontin polypeptide
  • Novel application of osteopontin polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the screening experiment of liver cancer marker

[0036] 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 ...

Embodiment 2

[0043] Embodiment 2, reverse protein chip operation steps

[0044] In this embodiment, the reverse protein chip technology is used to compare and analyze the 6 factors obtained in Example 1. The operation steps of the reverse protein chip are as follows:

[0045] Table 3, the reagent that the present embodiment adopts

[0046]

[0047]

[0048] (1) Sample processing: Serum samples provided by the experimenter were treated with appropriate buffer and serial dilution.

[0049] (2) Standard product preparation: prepare 6 factor recombinant proteins (see Table 3) to a concentration of 100g / ml (stock solution), dilute 5 times according to the initial concentration of each factor standard curve according to different 1 / 3 times, and set a blank control.

[0050] (3) Membrane preparation: the serum sample diluted in step (1), the gradient dilution standard prepared in step (2), positive control and blank control are spotted on the membrane; wherein, 800cw labeled streptavidi...

Embodiment 3

[0058] Embodiment 3, the preparation of stable OPN standard substance polypeptide

[0059] OPN is a protein composed of 314 amino acids, as shown in SEQ ID No.1 as follows:

[0060] MRIAVICFCL LGITCAIPVK QADSGSSEEK QLYNKYPDAV ATWLNPDPSQKQNLL APQNAVSSEETNDFKQETLPS KSNE SHDHMDDMDD EDDDDHVDSQ DSIDSNDSDD VDDTDDSHQSDESHHSDESD ELVTDFPTDL PATEVFTPVV PTVDTYDGRG DSVVYGLRSK SKKFRRPDIQ YPDATDEDITSHMESEELNG AYKAIPVAQD LNAPSDWDSR GKDSYETSQL DDQSAETHSH KQSRLYKRKA NDESNEHSDVIDSQELSKVS REFHSHEFHS HEDMLVVDPK KEEDKHLKFRI SHELDSAS SEVN

[0061] After a lot of research, the present invention finds that when the OPN recombinant protein is used as the calibrator to detect OPN in the sample, the stability of the standard product is worse than that of the natural protein due to the lack of glycosylation of the prokaryotic recombinantly expressed protein. Moreover, insoluble inclusion bodies are obtained during the purification of recombinant proteins, which are not easy to bind to specific antibo...

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PUM

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Abstract

The invention provides novel application of an osteopontin polypeptide, and particularly relates to application of the OPN polypeptide in preparation of a kit for quantitatively detecting a liver cancer marker. The inventor of the invention finds that the content change of the OPN has obvious correlation to the state of the liver cancer through long-term research and a large number of tests, the OPN is used as a marker, and the kit for detecting the content of the OPN is prepared by using the OPN, so that the existence, staging and metastasis of the liver cancer can be accurately diagnosed inan assisted manner.

Description

technical field [0001] The invention relates to the field of diagnosis of liver cancer, and specifically relates to a new application of osteopontin polypeptide. 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. Since tumor markers either do not exist in normal adult tissues...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574C07K14/52
CPCG01N33/6893G01N33/57484G01N33/57438C07K14/52
Inventor 黄若磐匡治州黄伟毛应清
Owner RAYBIOTECH INC GUANGZHOU
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