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

Immune globulin G4 detection kit and preparation method thereof

An immunoglobulin and detection kit technology, applied in the field of immunoglobulin G4 detection kit and its preparation, can solve the problems of HOOK effect, low sensitivity, slow detection speed, etc., to prevent HOOK effect, improve sensitivity, The effect of high coupling efficiency

Pending Publication Date: 2022-08-05
SHANGHAI KEHUA BIO ENG
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide an immunoglobulin detection kit and its preparation method, which mainly solve the technical problems in the prior art such as slow detection speed, low sensitivity, poor repeatability, and HOOK effect.

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
  • Immune globulin G4 detection kit and preparation method thereof
  • Immune globulin G4 detection kit and preparation method thereof
  • Immune globulin G4 detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 is the preparation process and using method of immunoglobulin G4 detection kit:

[0037] 1. Preparation of R1 reagent

[0038] Take 12g of 3-(N-morpholino)propanesulfonic acid (MOPS) in purified water, then add 5g of NaCl and 5g of NaN 3 , after stirring evenly, adjust the pH to 7.0, then add 5g of Tween-20, stir to dissolve, and set the volume to 1L as the first reagent R1;

[0039] 2. Preparation of R2 reagent

[0040] (1) Preparation of latex microsphere dilution:

[0041] Dissolve 12g 3-(N-morpholino)propanesulfonic acid (MOPS) in purified water, then add 10g NaCl, 50g sucrose, 50g trehalose and 5g NaN 3 , after stirring evenly, adjust the pH to 7.0, then add 10g of Triton X-405, stir and dissolve, and set the volume to 1L;

[0042] (2) Preparation of latex microsphere-carrier molecule complexes:

[0043] Take 1ml of carboxyl polystyrene latex microspheres (Bangs company) with a particle size of 60nm and dissolve it in 10ml of 25mmol / L 2-(N-morphol...

Embodiment 2

[0061] Embodiment 2 is the verification that carrier molecule improves reagent sensitivity:

[0062] In order to verify that the introduction of carrier molecules in the latex preparation process of the kit of the present invention improves the sensitivity of the reagent, four groups of kits are set for comparison.

[0063] Kit 1: the kit prepared in Example 1 of the present invention.

[0064] Kit 2: The difference from kit 1 is that the second reagent R2 in kit 2, in the process of latex-antibody coupling, the reduced antibody is directly coupled to latex microspheres without adding carrier molecules, and the rest of the preparation methods Same as Kit 1.

[0065] Kit 3: Immunoglobulin G Subtype 4 Detection Kit (Latex-Enhanced Immunity Turbidimetry - Listed Product A)

[0066] Kit 4: Immunoglobulin G4 Assay Kit (Latex-Enhanced Immunity Turbidimetry - Marketed Product B)

[0067] In this example, a fully automatic biochemical analyzer (Hitachi 7180) is used as the detectio...

Embodiment 3

[0087]Embodiment 3 is the verification that double particle size latex microspheres take into account both the sensitivity of reagents and the ability to resist HOOK:

[0088] In order to verify that the kit of the present invention adopts the double particle size latex microspheres to make the reagent take into account both the sensitivity and the ability of resisting the HOOK effect, five groups of kits were set up for comparison.

[0089] Serial dilutions (100%, 80%, 40%, 20%, 10%, 5%, 2.5%, 1.25%, 0.63%, 0.31%, 0.16%, 0.08%, 0.04%), three groups of kits were tested for gradient dilution samples at the same time, each sample was measured twice, and the mean value was calculated for comparison.

[0090] Kit 1: the kit prepared in Example 1 of the present invention.

[0091] Kit 5: The difference from kit 1 is that the second reagent R2 in kit 2 is coupled with latex with small particle size, and the rest of the preparation method is the same as kit 1.

[0092] Kit 6: The d...

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
particle diameteraaaaaaaaaa
particle diameteraaaaaaaaaa
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides an immunoglobulin G4 detection kit and a preparation method thereof. The immunoglobulin G4 detection kit is composed of a first reagent R1 and a second reagent R2, wherein the first reagent R1 comprises a zwitterionic buffer solution with the concentration of 25-200 mmol / L, an inorganic salt with the concentration of 5-10 g / L, a surfactant 1 with the concentration of 0.5-5 w% and a preservative with the concentration of 0.05-1 w%; the second reagent R2 comprises a zwitterionic buffer solution with the concentration of 25-200mmol / L, inorganic salt with the concentration of 5-10g / L, cane sugar with the concentration of 50-100g / L, trehalose with the concentration of 50-100g / L, 0.5-5w% of a surfactant 2, 1w%-5w% of latex microspheres which are marked by carrier molecules and are coupled with immune globulin G4 monoclonal antibodies, 1w%-5w% of a sealing agent and 0.05-1w% of a preservative. The kit has the advantages of high coupling efficiency, high sensitivity, hook effect resistance and the like.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis, and particularly relates to an immunoglobulin G4 detection kit and a preparation method thereof. Background technique [0002] Human immunoglobulin G4 is a subtype of immunoglobulin G, accounting for 4%-5% of total immunoglobulin G. Compared with other immunoglobulin G subtypes, the concentration of immunoglobulin G4 is lower. However, increased or decreased immunoglobulin G4 can be seen in a variety of immune system diseases. Immunoglobulin G4-related diseases are a new type of chronic, progressive inflammation with fibrosis and sclerosis of unknown etiology newly discovered in recent years, mainly characterized by significantly increased serum immunoglobulin G4 levels; Immunoglobulin G4-positive plasma cells infiltrate with fibrosis resulting in swollen or nodular / sclerotic lesions. The Chinese expert consensus on the diagnosis and treatment of immunoglobulin G4-related diseases (2...

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): G01N33/68G01N33/577G01N33/546G01N33/531
CPCG01N33/6854G01N33/577G01N33/54346G01N33/531
Inventor 王琦史小娟肖禄生彭珊
Owner SHANGHAI KEHUA BIO ENG
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