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Polypeptide marker and application thereof

A marker and peptide technology, applied in specific peptides, biological testing, biological material analysis, etc., can solve problems such as difficult reflection and short half-life of serum albumin, and achieve the effect of making up for low specificity

Active Publication Date: 2021-08-06
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] Based on this idea, Zhang Mei et al. took serum albumin as the research object and found three glucose-sensitive peptides (Zhang, M.; Xu, W.; Deng, Y.L., A New Strategy for Early Diagnosis of Type2Diabetes by Standard-Free, Label-Free LC-MS / MS Quantification of GlycatedPeptides. Diabetes 2013, 62(11), 3936-3942.), but the half-life of serum albumin in the human body is short, only about 20 days, difficult to reflect

Method used

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  • Polypeptide marker and application thereof
  • Polypeptide marker and application thereof
  • Polypeptide marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The establishment and detection analysis of embodiment 1 in vitro incubation diabetes model

[0047] 1. Preparation of In Vitro Glycosylation Samples

[0048] (1) The preparation of in vitro glycosylated samples refers to the methods in the literature (Tang Xiaojun, Lu Xian'e, Li Ge, et al. Analysis of risk factors for type 2 diabetes [J]. Chinese Public Health, 2004, 1(5): 11-15) , take 30mg of human hemoglobin standard product, add 10ml of PBS buffer solution with a concentration of 10mM to prepare a human hemoglobin standard product solution, since hemoglobin has 58 theoretical glycosylation sites, convert the molar ratio according to the physiological concentration of D-glucose and human hemoglobin , the molar ratio of glucose and human hemoglobin was designed according to the 5-fold excess of glucose.

[0049] (2) Mix 0.5ml of human hemoglobin standard solution with a concentration of 3mg / ml and 0.5ml of D-glucose solution with a concentration of 15mg / ml to prepar...

Embodiment 2

[0099] The verification of embodiment 2 clinical samples

[0100] 1. Collection of Clinical Samples

[0101] Diabetes mellitus is a group of endocrine and metabolic diseases mainly characterized by chronic hyperglycemia, clinically divided into three types: type 1, type 2 and gestational diabetes, of which type 2 diabetes is the most common. At present, there is no unified conclusion on the specific pathogenesis of various types of diabetes, but studies have shown that diabetes is the result of the joint action of genetic factors, autoimmune system factors and environmental factors. Among them, type 1 diabetes (T1DM) is mainly related to heredity, and insulin secretion is insufficient due to autoimmune deficiency of pancreatic islet cells. Patients mainly rely on insulin injections to compensate for the lack of insulin secretion. Type 2 diabetes (T2DM) is a chronic progressive metabolic disease. The incidence of T2DM is high, and its complications are harmful. If T2DM is n...

Embodiment 4

[0124] Example 4 Potential Application of Polypeptide Markers in Diabetes Diagnosis

[0125] 1. Synthesis of stable isotope-labeled marker peptides (C 13 mark or n 15 tags are fine)

[0126] According to the internal standard peptide and sensitive peptide obtained by screening verification in Example 1 and Example 2, design and synthesize N based on solid-phase synthesis method 15 Labeled internal standard peptide and N 15 Labeled sensitive peptide, in addition to relative molecular mass, N 15 Labeled internal standard peptide and internal standard peptide, N 15 There is almost no difference in the properties of the labeled sensitive peptide and the sensitive peptide. In the kit, N 15 Labeled internal standard peptide and N 15 The amount of substances such as labeled sensitive peptides were mixed to obtain N 15 Standard peptides were stored at -80°C for future use.

[0127] 2. Pretreatment of clinical samples

[0128] The patient's whole blood sample was selected for ...

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Abstract

The invention relates to a polypeptide marker and application thereof, and belongs to the technical field of biology. An amino acid sequence of the polypeptide marker is SEQ ID No.1 in an amino acid sequence table. The invention further provides a polypeptide marker with an amino acid sequence being SEQ ID No.2 in the amino acid sequence table. The application refers to application of the polypeptide marker to preparation of a kit for early diagnosis of diabetes mellitus. According to the invention, a quantitative peptide biomarker is provided as a strategy for clinical early diagnosis of the diabetes mellitus, related indexes for preventing and monitoring the diabetes mellitus are obtained, and the ratio of peak areas or peak intensities of peptide fragments obtained by liquid chromatography-mass spectrometry of sensitive peptides and internal standard peptide fragments is adopted as a glycosylation modification degree index, so that the problems of relatively large individual difference between clinical samples and relatively high difficulty in relative quantification, which is of great significance to early diagnosis and effective intervention of the diabetes mellitus.

Description

technical field [0001] The invention relates to a polypeptide marker and its application, in particular to a polypeptide marker for detecting early diabetes, which belongs to the field of biotechnology. Background technique [0002] Diabetes Mellitus (DM) is a metabolic disease caused by various etiologies and pathogenesis, relative or absolute insufficiency of insulin in the body, which leads to disorders of sugar, protein and fat metabolism in the body, and hyperglycemia as the main clinical manifestation. . 90% of diabetic patients worldwide suffer from type 2 diabetes (Type 2Diabetes, T2D), which is a chronic progressive metabolic disease. In the early stage of type 2 diabetes, the body maintains a normal level of blood sugar by increasing the secretion of insulin, and the β-islet cells are in an overloaded working state. If things go on like this, it will lead to damage to the islet cells, forming a "high blood sugar concentration-islet cell damage" - a vicious circle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/805G01N33/68G01N30/02
CPCC07K14/805G01N33/6893G01N30/02G01N2333/805G01N2800/042
Inventor 张永谦信树辰洪杰邓玉林
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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