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Method for establishing quantitative reference range of healthy human urine proteome and database of healthy human urine proteome

A technology of proteome and reference range, 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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  • 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 individuals, rather than proteins that truly reflect the differences between disease and control states. It is the main reason why no new urinary protein markers have been successfully applied in clinical practice through proteomics methods

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  • Method for establishing quantitative reference range of healthy human urine proteome and database of healthy human urine proteome
  • Method for establishing quantitative reference range of healthy human urine proteome and database of healthy human urine proteome
  • Method for establishing quantitative reference range of healthy human urine proteome and database of healthy human urine proteome

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preparation example Construction

[0031] 1. Preparation of urine protein samples

[0032] For the collected healthy human urine samples, the invention adopts the following methods based on ultracentrifugation and reduction to obtain urine protein samples:

[0033] (1) Centrifuge 10ml of urine sample at 4°C for 20 minutes with a centrifugal force of 100,000g, discard the supernatant and keep the precipitate;

[0034](2) Transfer the above precipitate to a centrifuge tube, add 60 μl of resuspension buffer (50 mM Tris, 250 mM sucrose, pH 8.5) into the centrifuge tube, let it stand at room temperature for 10 minutes, and blow the resuspended precipitate fully with a pipette ;

[0035] (3) Add dithiothreitol to the above resuspended pellet to a final concentration of 50 mM, and heat at 80°C for 10 minutes to remove most of the uromodulin protein in the sample;

[0036] (4) Add washing buffer (10mM triethanolamine, 100mM sodium chloride, pH7.4) to 400ul, then centrifuge for 20 minutes under 4 conditions with a c...

Embodiment 1

[0058] Example 1. Establishing a data set for evaluating physiological fluctuations and differences in the urine proteome of healthy individuals, and evaluating the physiological fluctuations in the urine proteome individual

[0059] The process of building a dataset includes:

[0060] 1) Sampling: Continuously collect urine samples from 17 informed consent volunteers with different time spans, see Table 1 for sampling time and quantity;

[0061] 2) Preparation of urine protein samples: each urine sample collected was made into a urine protein sample according to the above-mentioned method, and each urine sample was made into a urine protein sample (peptide sample containing 2 components));

[0062]3) Detection: each urine protein sample was detected according to the above-mentioned method 2, and the mass spectrometry data of each urine protein sample was obtained, and the first row U001-1 in Table 1 (one of the samples collected by volunteer U001 in 24 hours Urine protein sa...

Embodiment 2

[0088] Example 2. Establishing a data set for assessing physiological fluctuations and differences between individuals in the urine proteome of healthy people, and evaluating the physiological fluctuations in the urine proteome among individuals

[0089] The data collection of healthy human urine proteome is the same as that in Example 1.

[0090] In this embodiment, the sub-dataset BPRC (sub-dataset 2, see Table 5) composed of 178 urine proteome data of 150 volunteers was collected.

[0091] Table 5. 178 urine proteome data sub-datasets BPRC including 150 healthy volunteers

[0092]

[0093] Subdataset 2 (BPRC) and subdataset 1 (BCM) were merged to obtain a total dataset including 497 urine proteome data of 167 healthy volunteers (integrated Table 4 and Table 5, omitted here). The total dataset can also be divided into male and female urinary proteome sub-datasets according to the sex of the volunteers. Sub-dataset 1 (including 319 urine proteome data of 17 healthy volun...

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Abstract

The invention discloses a method for establishing a quantitative reference range of a healthy human urine proteome and a database of a healthy human urine proteome. The method comprises the steps thata statistical quantity of healthy human urine samples are made into urine protein samples, protein species and quantification of each protein are determined by mass spectrometric detection, library searching and quantification to form urine proteome data, different urine proteome data is collected into different urine proteome sub-data sets and total data sets, and the quantitative reference range of the healthy human urine proteome is obtained by the data calculation of the data sets. The established quantitative reference range of the human urine protein in the healthy human urine proteomedatabase can better exclude interference from physiological fluctuations and inter-individual differential protein in the discovery of urine protein biomarkers.

Description

technical field [0001] The present invention belongs to the establishment of biomarker data in the field of medical biology, and in particular relates to a method for establishing a quantitative reference range for urine proteome of healthy humans by using a healthy human urine proteome data set that can cover physiological fluctuations and differences within and between individuals and the resulting method. The established healthy human urine proteome database. Background technique [0002] Urine is the most commonly used body fluid sample except blood in clinical tests. The detection of bilirubin, glucose, ketone bodies, protein, blood cells and other indicators in urine routine is used for the diagnosis or efficacy monitoring of various diseases. In view of the important value of urine testing in health medicine, scientists from all over the world have been using proteomics technology to try to find new protein markers from urine for disease diagnosis, prognosis judgment,...

Claims

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Application Information

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IPC IPC(8): G06F19/28
CPCG16B50/00
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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