Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Detection kit for anti-peptidyl prolyl cis-trans isomerase D-IgG antibody

A peptidyl prolyl cis, detection kit technology, applied in the field of biomedicine, can solve problems such as blanks and unreported relationships, and achieve the effects of easy operation, easy cleaning and separation, and improved reaction speed

Active Publication Date: 2021-09-28
ZHEJIANG UNIV
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is no report on the relationship between peptidyl-prolyl cis-trans isomerase D and nephrotic syndrome
In the prior art, there is currently no research on the identification of autoimmune nephrotic syndrome by detecting serum anti-peptidyl-prolyl cis-trans isomerase D-IgG antibodies

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Detection kit for anti-peptidyl prolyl cis-trans isomerase D-IgG antibody
  • Detection kit for anti-peptidyl prolyl cis-trans isomerase D-IgG antibody
  • Detection kit for anti-peptidyl prolyl cis-trans isomerase D-IgG antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Peptidyl-prolyl cis-trans isomerase D on podocytes is the target antigen for autoantibodies in patients with autoimmune nephrotic syndrome

[0037] (1) Extraction of total protein of glomerular podocytes: culture podocyte strain (MPC5), wash 2-3 times with PBS, and then use a focused ultrasonic instrument (Covaris S220, Gene) in a medium containing 30mm Tris-HCl, 8m urea, 4% CHAPS and protease inhibitors (#ab65621; Abcam, 1:200 dilution) were fully lysed on ice in the lysis buffer, and then the sample was placed in a centrifuge at 12000g, 4°C for 30min. The supernatant was collected, which was the collected glomerular podocyte total protein. The total protein concentration of collected glomerular podocytes was determined by BCA protein concentration assay kit. (2) Two-dimensional electrophoresis: the total protein of glomerular podocytes was extracted for two-dimensional electrophoresis, then transferred to a nitrocellulose membrane, incubated with serum from...

Embodiment 2

[0038] Example 2 Expression and purification of recombinant peptidyl-prolyl cis-trans isomerase D antigen protein

[0039] The method of genetic engineering is used to use the gene encoding peptidyl-prolyl cis-trans isomerase D protein as a template to carry out PCR amplification, and then construct an expression vector for protein expression. The antigenic protein expressed in the present invention contains the tag peptide of the His tag. The expressed recombinant protein was purified by nickel column affinity chromatography, ion affinity chromatography, hydrophobic column, molecular sieve, etc. Finally, the molecular weight of the recombinant protein peptidyl-prolyl cis-trans isomerase D was identified by SDS-PAGE as 41KDa, the results are shown in figure 2 .

Embodiment 3

[0040]Example 3 The present invention uses an orthogonal experiment design to optimize the reaction conditions of the kit According to the coating concentration of antigen peptidyl-prolyl cis-trans isomerase D (50 μg / mL, 100 μg / mL, 150 μg / mL, 200 μg / mL four coating Concentration), each reaction time (30min, 45min) and temperature (25°C, 35°C), the optimal dilution of the enzyme-labeled secondary antibody (1:100, 1:500, 1:1000, 1:1500 four dilutions) Choose an orthogonal table for 4 factors, each factor is repeated at 2 levels for standard positive serum and standard negative serum. Select the ratio (P / N) of the highest luminescence value (P) of the positive serum to the lowest luminescence value (N) of the negative serum. The average P / N value of repeated determinations was determined through statistical processing to determine the optimal coating conditions and the optimal dilution of the secondary antibody for orthogonal optimization conditions, which significantly improved ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides a detection kit for an anti-peptidyl prolyl cis-trans isomerase D-IgG antibody. A detected serum sample is in contact with target antigen peptidyl-prolyl cis-trans isomerase D polypeptide or a binding fragment of the isomerase D polypeptide, an immune reaction is carried out, an antigen-antibody compound is formed, and the anti-peptidyl prolyl cis-trans isomerase D-IgG autoantibody is detected through the formed antigen-antibody compound. The D-IgG antibody aiming at the anti-peptidyl prolyl cis-trans isomerase is identified in the nephrotic syndrome of children for the first time, and the detection kit is provided aiming at the antibody. A good detection method is provided for identifying the autoimmune nephrotic syndrome related to the anti-peptidyl prolyl cis-trans isomerase D-IgG antibody at home and abroad.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a detection kit for anti-peptidyl prolyl cis-trans isomerase D-IgG antibody. Background technique [0002] Primary nephrotic syndrome (PNS) is caused by a variety of factors that increase the permeability of the glomerular basement membrane, and then increase the filtration of plasma protein and lose it in the urine, which leads to a series of pathological changes. clinical syndrome. PNS is a common glomerular disease in children, with an annual incidence of 1.15-16.9 / 100000 according to reports. The pathological type is minimal change nephrotic syndrome (MCNS), which is the most common. According to the response effect of PNS to hormone therapy, it can be divided into steroid-sensitive type (SSNS), steroid-dependent type (SDNS), and steroid-dependent type (SDNS). Drug-resistant type (SRNS). Although the vast majority of children are sensitive to hormone therapy and have a go...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/564G01N33/573G01N33/543G01N21/76
CPCG01N33/564G01N33/573G01N33/54326G01N21/76G01N2333/99G01N2800/24G01N2800/347
Inventor 叶青毛建华张俊峰
Owner ZHEJIANG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com