Method for obtaining tumor urine protein marker and obtained stray urine protein library related to tumor

A tumor-related, urinary protein technology, applied in the field of medicine and biology, to achieve the effect of eliminating interference

Active Publication Date: 2018-07-27
北京松果天目健康管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of high-throughput deep quantitative urine proteome detection methods, the candidate markers found through small sample sizes in the discovery stage are usually proteins that differ between different individua...

Method used

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  • Method for obtaining tumor urine protein marker and obtained stray urine protein library related to tumor
  • Method for obtaining tumor urine protein marker and obtained stray urine protein library related to tumor
  • Method for obtaining tumor urine protein marker and obtained stray urine protein library related to tumor

Examples

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preparation example Construction

[0053] 1. Preparation of urine protein sample

[0054] The invention uses the following methods based on ultracentrifugation and reduction to obtain urine protein samples for the collected urine samples of healthy people and tumor patients:

[0055] (1) A 10ml urine sample was centrifuged at 4°C for 20 minutes at a centrifugal force of 100000g, the supernatant was discarded, and the precipitate was retained;

[0056] (2) Transfer the above-mentioned pellet to a centrifuge tube, add 60μl of resuspension buffer (50mM Tris, 250mM sucrose, pH8.5) to the centrifuge tube, let stand at room temperature for 10 minutes, and pipette to resuspend the pellet ;

[0057] (3) Add dithiothreitol to the above resuspended precipitate to a final concentration of 50mM, and heat at 80°C for 10 minutes to remove most of the uromodulin in the sample;

[0058] (4) Add washing buffer (10 mM triethanolamine, 100 mM sodium chloride, pH 7.4) to 400 ul, and then centrifuge at 4 at a centrifugal force of 100,000 f...

Embodiment 1

[0091] Example 1. Establish a data set for evaluating the physiological fluctuations and differences in the urine proteome of healthy people, and evaluate the physiological fluctuations in the urine proteome of individuals

[0092] The process of establishing a data set includes:

[0093] 1) Sampling: Continuously collect urine samples of 17 informed consent volunteers with different time spans. See Table 1 for sampling time and quantity;

[0094] 2) Preparation of urine protein samples: each collected urine sample is made into a urine protein sample according to the aforementioned method, and each urine sample is made into a urine protein sample (a peptide sample containing 2 components));

[0095] 3) Detection: Test each urine protein sample according to the above two methods, and obtain the mass spectrum data of each urine protein sample. Take U001-1 in the first row of Table 1 (one of the 24 hours collected by volunteer U001) Urine protein sample made from urine sample) as an exam...

Embodiment 2

[0120] Example 2. Establish a data set for evaluating the physiological fluctuations and differences between individuals in the urine proteome of healthy people, and evaluate the physiological fluctuations between individuals in the urine proteome

[0121] The data collection of healthy human urine proteome is the same as in Example 1.

[0122] In this example, a sub-data set BPRC composed of 178 urine proteomic data of 150 volunteers was collected (see Table 5).

[0123] Table 5. 178 urine proteomic data subdataset BPRC including 150 healthy volunteers

[0124]

[0125]

[0126] The sub-data set BPRC and the sub-data set BCM were merged to obtain 497 urine proteome data set A including 167 healthy volunteers (integrated table 4 and table 5, omitted here). Data set A can also be divided into male and female urine proteome sub-data sets according to the sex of the volunteers. The sub-data set A1 is composed of 350 healthy human urine proteome data to establish a quantitative reference ...

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Abstract

The invention provides a method for obtaining a tumor urine protein marker and an obtained stray urine protein library related to a tumor. The method comprises the steps that based on a built quantitative reference range of human urine protein in a healthy human urine protein database, a mode of hypergeometric distribution detection is used for screening stray protein as the tumor urine protein marker from a urine proteome dataset of a tumor patient, and the stray urine protein library related to the tumor is established. The method for obtaining the tumor urine protein marker and the obtainedstray urine protein library related to the tumor can better eliminate the interference of physiological fluctuation and inter-individual differential protein in the process of finding a urine proteinbiomarker.

Description

Technical field [0001] The invention belongs to the establishment of biomarker data in the field of medical biology, and particularly relates to screening the urine proteome of tumor patients using healthy human urine proteome quantitative reference range data to obtain tumor urine protein markers (ie outlier urine protein) The method and the establishment of tumor-related outlier urine protein library. Background technique [0002] Urine is the most commonly used body fluid sample other than blood in clinical tests. The detection of bilirubin, glucose, ketone bodies, protein, blood cells and other indicators in urine routines is used to diagnose or monitor the efficacy of various diseases. In view of the important value of urine testing in health medicine, scientists around the world have been using proteomics technology to try to find new protein markers in urine for disease diagnosis, prognosis determination, and efficacy testing. The current research and development process ...

Claims

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Application Information

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IPC IPC(8): G06F19/18G01N30/02
CPCG01N30/02G16B20/00
Inventor 秦钧甄蓓冷文川倪晓天路天元王广舜孙长青钟博文
Owner 北京松果天目健康管理有限公司
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