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Kit for detecting anti-peroxiredoxin-1-IgG antibody

A peroxide, -1-igg technology, applied in the field of biomedicine, can solve the problems of peroxiredoxin-1 expression and the existence of peroxiredoxin-1 that have not been reported, so as to increase the coating surface area, reduce the probability of cross-infection, and simple operation Effect

Active Publication Date: 2021-09-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the expression of peroxiredoxin-1 and the presence of autoantibodies to peroxiredoxin-1 have not been reported in nephrotic syndrome
In addition, in the prior art, there is no application based on the target peroxiredoxin-1 or its autoantibody as a serological marker in autoimmune nephrotic syndrome
Identification of autoimmune nephrotic syndrome by detection of serum anti-peroxiredoxin-1-IgG antibodies is blank

Method used

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  • Kit for detecting anti-peroxiredoxin-1-IgG antibody
  • Kit for detecting anti-peroxiredoxin-1-IgG antibody
  • Kit for detecting anti-peroxiredoxin-1-IgG antibody

Examples

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Embodiment 1

[0035] Example 1 Peroxiredoxin-1 protein on foot cells is a target antigen for autoimmunity antibodies in autoimmune nephropathy syndrome

[0036] The present invention has been studied by a large number of clinical and molecular mechanisms in the previous period. The serum IgG levels in patients with nephropathy syndrome were highly discovered, and it was confirmed that Peroxiredoxin-1 on foot cells was the target antigen against autoimmunity in patients with autoimmune nephropathy. Specifically, the following (1) Extraction of total protein of glomerular foot cells is specifically implemented: a culture of foot cell strain (MPC5) was cultured, 2-3 times with PBS, and then focused ultrasound instrument (Covaris S220, Gene) containing 30 mm Tris -HCl, 8M urea, 4% CHAPS and protease inhibitor (# AB65621; ABCAM, 1: 200 diluted) lytic solution, then place the sample in centrifuge, 12000 g, 4 ° C, centrifuge for 30 min . The supernatant is collected, that is, the total protein collect...

Embodiment 2

[0037] Example 2 recombinant antigenic peroxiredoxin-1 expression and purification

[0038] Using the genetic engineering method to encode Peroxiredoxin-1 protein by a template, PCR amplification is performed, and then the expression vector is used for protein expression. The antigenic protein of the present invention contains a tag peptide of the HIS label. Expressive recombinant protein is purified by nickel pillars, ion affinity chromatography, hydrophobic columns, molecular sieves, and the like, and finally use SDS-PAGE to identify the molecular weight of recombinant protein peroxiredoxin-1. figure 2 .

Embodiment 3

[0039] Example 3 The present invention optimizes the reactive conditions of the kit with orthogonal test design.

[0040] According to the antigen Peroxiredoxin-1 pack of concentrations (50 μg, 80 μg, 100 μg, 150 μg of four packs), each reaction time (15 min, 30 min, 45min) and temperature (25 ° C, 37 ° C), and the optimal dilution of the enzyme-standard secondary antibodies 4 factors (1: 100, 1: 500, 1: 1: 1: 1500 four dilution) are selected in the formation, each factor, repeatedly determined standard positive serum and standard negative serum at 2 levels. The ratio (P / N) of the highest optical signal value (P) of positive serum and the minimum optical signal value (N) of the negative serum, the highest optical signal value (P) of positive serum, and the lowest optical signal value of the negative serum ( N) The ratio (P / N). Through orthogonal design, we got this kit for optimal antigen Peroxiredoxin-1 pack of concentrations of 80 μg / ml, solid phase film immunoassay anti-P...

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Abstract

The invention provides a kit for detecting an anti-peroxiredoxin-1-IgG antibody. The kit consists of antigen protein peroxiredoxin-1, a solid-phase carrier, a labeled antibody, an antigen diluent, a sample dilution buffer solution, an antibody diluent, a substrate color developing agent, a washing solution, a standard substance, a positive quality control substance and a negative quality control substance. According to the kit disclosed by the invention, the anti-peroxiredoxin-1-IgG antibody in serum to be detected is detected by utilizing an indirect method reaction principle in combination with magnetic particle chemiluminescence immunoassay. According to the invention, the autoantibody aiming at the target antigen peroxiredoxin-1 in the serum of the patient with the autoimmune nephrotic syndrome is identified for the first time; the kit provided by the invention provides a basis for molecular mechanism research and clinical diagnosis and treatment of autoimmune nephrotic syndromes related to peroxiredoxin-1 and peroxiredoxin-1-IgG autoantibodies at home and abroad.

Description

Technical field [0001] The present invention belongs to the field of biomedical technology, and involves a kit for detecting an anti-oxide reductase-1-IgG antibody. Background technique [0002] In recent years, there have been more and more types of kidney disease in recent years, including the highest incidence of autoimmune nephropathy syndrome, seriously harmful children's physical and mental health. Autographic immune nephropathy syndrome, because the permeability of glomerular filtration membrane increases, plasma protein filtration increases, causing a large number of protein urine, and thus causes patients mainly manifesting a large number of protein urine, low proteinmia, Clinical syndrome such as high edema and other characteristics. Scholars such as Ali have been observed that after transplantation of the kidneys from the patients with difficulty microaceous nephropathy syndrome, the renal function did not have any protein urine, which showed that the microxidative nep...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/573G01N33/543
CPCG01N33/6854G01N33/6893G01N33/58G01N33/573G01N33/54326G01N2333/908G01N2800/347C12N9/0065C12Y111/01015
Inventor 叶青毛建华田丹丹
Owner ZHEJIANG UNIV
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