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Use of serine with mass shift at position 208 of akap3 protein in the preparation of diagnostic reagents for severe oligospermia and asthenospermia

Technology of a diagnostic reagent, serine, used in the field of medicine and molecular diagnostics

Active Publication Date: 2018-03-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports on non-coding amino acids as biomarkers associated with severe oligoasthenospermia

Method used

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  • Use of serine with mass shift at position 208 of akap3 protein in the preparation of diagnostic reagents for severe oligospermia and asthenospermia
  • Use of serine with mass shift at position 208 of akap3 protein in the preparation of diagnostic reagents for severe oligospermia and asthenospermia
  • Use of serine with mass shift at position 208 of akap3 protein in the preparation of diagnostic reagents for severe oligospermia and asthenospermia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Example 1: Screening of biomarkers associated with severe oligoasthenospermia

[0125] The specific screening method is as follows:

[0126] 1. Sample processing and experimental analysis

[0127] 1. Extraction of the whole protein of sperm cells: The same amount of severe oligospermia and normal sperm samples were washed three times with DPBS, and the same amount of RIPA lysate was added to sonicate for 1-2 minutes, incubated on ice for 30 minutes to lyse, and centrifuged at 14,000g at 4°C ×20min to take the supernatant. Protein concentration was determined using the Bradford method.

[0128]2. Proteolysis: Take approximately 150 μg of sperm protein from the samples of severe oligospermia and normal sperm, use 10% polyacrylamide gel electrophoresis (SDS-PAGE) to separate the proteins, and divide each into 5 parts for enzymatic hydrolysis. Peptides were desalted using ziptip.

[0129] 3. Mass Spectrometry: Nanoflow Liquid Chromatography Separation: Phase A: Water co...

Embodiment 2

[0185] Embodiment 2: clinical detection verification

[0186] Taking 4 healthy samples and 8 clinically diagnosed severe oligoasthenospermia samples as the research objects for verification, the serine at the 208th position of the AKAP3 protein and the mass deviation of -113.05347 at the 186th position of the AKAP4 protein were detected respectively in the above samples. Asparagine shifted, asparagine with mass shift of -114.04278 at position 186 of AKAP4 protein, glutamine with mass shift of -17.02660 at position 617 of AKAP4 protein, lysine with mass shift of +211.09682 at position 733 of AKAP4 protein , lysine with +42.01108 mass shift at position 531 of ATP5A1 protein, lysine with +42.01108 mass shift at position 87 of COX4I1 protein, threonine with +79.96685 mass shift at position 64 of GAPDHS protein and 692 position of KIAA1683 protein The respective detection frequencies of serine with +79.96685 mass shift occurred, and the samples to be tested were diagnosed according...

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Abstract

The invention discloses application of serine, which is used as a biological marker and has +79.96685 mass shift at a 208 site of AKAP3 (A-kinase Anchor Protein 3), to preparation of a diagnostic reagent or a treatment medicine of severe oligoasthenospermia. According to the application disclosed by the invention, the severe oligoasthenospermi can be diagnosed through testing frequency of the +79.96685 mass shift occurring at the 208 site of the AKAP3, and new diagnosis and new treatment target spots are provided for the severe oligoasthenospermia.

Description

technical field [0001] The invention relates to the technical fields of medicine and molecular diagnosis, in particular to the use of a serine with a mass shift at position 208 of the AKAP3 protein as a biomarker in the preparation of a diagnostic reagent for severe oligospermia and weak sperm. Background technique [0002] Infertility has become a worldwide reproductive health problem, of which the infertility caused by male factors accounts for about 50%, and it is on the rise in recent years. The main cause of male infertility is oligozoospermia. According to the World Health Organization standard, if the number of a-class sperm is less than 25%, the number of (a+b)-class sperm is less than 50%, and the sperm motility rate is lower than 60%, it can be diagnosed as asthenozoospermia. Oligospermia refers to a condition in which the number of sperm in semen is lower than that of normal fertile men. When a man's sperm is less than 20 million per milliliter, it is oligospermi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N33/68
CPCG01N30/02G01N33/6848G01N2800/367
Inventor 巩晶杨静华陈新骏陈子江
Owner SHANDONG UNIV
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