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

Assay method and assay kit for hepatitis b virus s antigen

A technology for hepatitis B virus and s antigen, which is applied in viral peptides, chemical instruments and methods, measuring devices, etc., can solve the problems of low measured values ​​and obstacles, and achieve the effect of reducing the impact

Pending Publication Date: 2020-07-14
FUJIREBIO CO LTD
View PDF7 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the case where the HBs antibody in the patient's blood is positive, when HbsAg is analyzed in the above-mentioned method using an antibody that binds to the common antigenic determinant a, the binding of the antibody used in the analysis method to HBsAg is subject to the patient's Obstruction of HBs antibody, the measurement value becomes lower

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
  • Assay method and assay kit for hepatitis b virus s antigen
  • Assay method and assay kit for hepatitis b virus s antigen
  • Assay method and assay kit for hepatitis b virus s antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093]

[0094] (1) Preparation of non-reducing and reducing HBsAg immobilized plates

[0095] Commercially available native HBsAg (Subtype ad, manufactured by TRINA) was diluted to 12.2 μg / mL with PBS or PBS containing 6M urea to prepare antigen dilutions (urea(-), urea(+)). Dithiothreitol (DTT) was diluted with ion-exchanged water to prepare reducing agent solutions of 1, 10, 50, 100, 200, 500, and 1000 mM, respectively. To each well of a 96-well microplate (manufactured by Nunc), 90 μL of the diluted antigen solution was dispensed per well, and 10 μL of the reducing agent solution was added thereto. After standing at room temperature for 60 minutes, PBS or PBS containing 6M urea was added thereto at 80 μL / well. After standing overnight at 4°C, wash with PBS 3 times. 350 μL of blocking solution (1% BSA, 3% sucrose, PBS) was dispensed, and after standing at room temperature for 3 hours, the blocking solution was removed by suction and air-dried at room temperature.

[00...

Embodiment 2

[0106]

[0107] For 8 cases of anti-HBs antibody (HBsAb) positive serum samples, the reactivity test to reduced antigen was carried out. Non-reducing and reducing antigen immobilized plates were prepared in the same manner as in Example 1.

[0108] The following samples were prepared.

[0109] a) Monoclonal antibody solution

[0110] Antibody A and D were diluted to 1 μg / mL with antibody diluent to prepare antibody solutions.

[0111] b) Autoantibody negative serum

[0112] Use 2 autoantibody-negative sera (N1, N2).

[0113] c) Autoantibody-positive serum

[0114] Eight cases of autoantibody-positive sera (P1-P8) were used. The antibody titer of each sample was measured using LUMIPULSE HBsAb-N (manufactured by Fuji Rebio). Table 3 shows the measured values ​​of the respective samples.

[0115] [table 3]

[0116]

[0117]

[0118] 100 μL / well of the antibody dilution was dispensed into each plate, and 10 μL of the antibody solution and the sample were added to e...

Embodiment 3

[0122]

[0123] Here it was investigated whether human anti-HBsAg antibodies contained in autoantibody-positive samples would compete with various monoclonal antibodies.

[0124] Serum samples with positive autoantibodies were used in 6 cases (P9-P14). The antibody titer of each serum sample was measured using LUMIPULSE HBsAb-N (manufactured by Fuji Rebio). The measured values ​​of each sample are shown in Table 3.

[0125]Non-reduced antigen immobilized plates were prepared in the same manner as in Example 1. Dilute antibodies A, D, E, and F to 100 μg / mL with antibody diluent, and dispense at 100 μL / well. Serum samples P9 to P14 were each diluted 5-fold with PBS, and 10 μL / well was further dispensed into the wells into which the antibody was dispensed. After standing at room temperature for 60 minutes, wash 5 times with plate washing solution. 100 μL / well of the labeled antibody solution (a solution obtained by diluting HRP-labeled anti-mouse IgG antibody or HRP-labeled...

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
diameteraaaaaaaaaa
Login to View More

Abstract

Disclosed are a high-sensitivity HBsAg assay method and assay kit that do not involve a strong acid or alkaline treatment during sample pretreatment and are not readily affected by autoantibodies. Anassay method for the hepatitis B virus s antigen in a sample that has been isolated from an organism, the method including: a pretreatment step for mixing the sample with a pretreatment reagent that includes a reducing agent to reduce the hepatitis B virus s antigen; and an immunoassay step for performing an immunoassay for hepatitis B virus s antigen on the pretreated sample using an antibody that can achieve an antigen-antibody interaction with a reduced peptide that comprises the 98th-179th amino acids of hepatitis B virus s antigen and / or an antigen-binding fragment of the antibody.

Description

technical field [0001] The invention relates to a detection method and a detection kit for hepatitis B virus s antigen. Background technique [0002] Hepatitis B is currently the most common disease among liver diseases, and it is a viral hepatitis caused by hepatitis B virus (HBV) infecting people. [0003] The main body of the hepatitis B virus is a particle called a "large spherical (Dane) particle" in the shape of a spherical particle having a double-layer structure with a diameter of 42 nm. It is known that the surface of a large spherical particle is covered with a surface antigen called hepatitis B virus s antigen (HBsAg), and further inside there is a core structure with a diameter of 27 nm, which contains HBc antigen (core antigen), HBe antigen, and Circular double-stranded DNA encoding viral genes. [0004] HBsAg is a tumor-constituting coat protein on the surface of infectious HBV particles, which is contained in a lipid bilayer membrane derived from hepatocytes...

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): G01N33/576C07K14/02
CPCC07K14/02G01N33/576G01N33/5764Y02A50/30G01N33/5306G01N33/563G01N2333/02
Inventor 大植千春饭田久美子青柳克己八木慎太郎
Owner FUJIREBIO CO LTD
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