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

Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof

A quantitative detection method, Golgi protein technology, applied in biological testing, material inspection products, etc., to achieve high sensitivity and specificity, shorten operation time, and improve sensitivity and specificity

Inactive Publication Date: 2011-03-23
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
View PDF5 Cites 33 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] According to the data retrieval carried out by the inventor, there are no other reports on the detection of GP73 double monoclonal antibody sandwich ELISA method at home and abroad

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
  • Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof
  • Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof
  • Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The present invention involves multiple optional monoclonal antibodies whose recognition sites can be seen figure 1 , figure 1 It is a schematic diagram of the GP73 protein structure of the present invention, and the monoclonal antibodies 1F2, 1A6, and 5F10 obtained by the inventors through screening recognize the IV region, and 5B12 and 9D2 recognize the V region.

[0042] In the sandwich ELISA quantitative detection method of Golgi protein GP73 of the present invention, three groups of paired monoclonal antibodies can be selected, namely 5B12-5F10; 5B12-1A6; 5F10-1A6. Select one of the pairing examples as follows, wherein, the coated antibody mAb 1 For 5B12, detection antibody mAb 2 It is HRP-labeled 5F10, namely HRP-5F10.

[0043] ①Use pH 9.6, 0.05mol / L carbonate buffer as the coating solution, dilute the coating antibody 5B12, the working concentration is 1ug / mL, add to 96-well microplate, 100μL / well, moisturize, 4 degrees overnight;

[0044] ②Wash the plate 3 ...

Embodiment 2

[0054] A preferred embodiment of the sandwich ELISA quantitative detection kit for Golgi protein GP73 of the present invention includes coating antibody and detection antibody, blocking solution, coating solution, diluent, chromogen, stop solution, washing solution and standard, including The target antibody and the detection antibody are two monoclonal antibodies against different epitopes in the extracellular region of GP73, and the standard product is the prokaryotic expressed and purified GP73 full-length protein. The detection antibody is a horseradish peroxidase (HRP)-labeled antibody, and the chromogenic reagent is 3,3'5,5'-tetramethylbenzidine (TMB) single-component chromogenic solution. The blocking solution is phosphate buffer containing 5% calf serum, 0.1% Tween20 (Tween20) and 0.01% preservative procline300. The coating antibody and the detection antibody are two monoclonal antibodies directed against different epitopes of the extracellular region of GP73, and the ...

Embodiment 3

[0065] Utilize the method described in embodiment 2, the GP73 content (SGP73) of serum sample is measured, and sample comprises healthy person (70 parts); Non-hepatic patient (131 parts); Hepatitis patient (57 parts); Liver cirrhosis patient ( 60); liver cancer patients (28). For specific results, see image 3 The SGP73 content of hepatitis patients was 110.83±60.33ng / mL, the SGP73 content of liver cirrhosis patients was 124.22±51.28ng / mL, the SGP73 content of liver cancer patients was 135.72±59.21ng / mL, and the SGP73 content of all liver disease patients was 121.18±56.9ng / mL ; The content of SGP73 in patients without liver disease was 62.80±34.91ng / mL, and the content of SGP73 in healthy people was 53.45±17.07ng / mL. Figure 4 is based on image 3 The histogram of the SGP73 content of different liver disease patients, non-hepatic disease patients and normal people, and the comparison of different groups. Figure 4 The data showed that the content of SGP73 in patients with h...

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

No PUM Login to View More

Abstract

The invention provides a sandwich enzyme-linked immuno sorbent assay (ELISA) quantitative detection method of Golgi protein GP73. Two strains of monoclonal antibodies aiming at different epitopes of a GP73 extracellular domain are used as a coated antibody and a detection antibody; and the GP73 content is detected through substrate developing and a standard curve. A sandwich ELISA quantitative detection kit of the GP73 comprises the coated antibody, the detection antibody, sealing solution, washing buffer, a color reagent, stop solution and a standard substance, wherein the coated antibody and the detection antibody are two strains of monoclonal antibodies aiming at different epitopes of the GP73 extracellular domain; and the amino acid sequences of proteins recognized by the monoclonal antibodies are shown as SEQ ID No. 1 and SEQ ID No. 2 respectively. Through the sandwich ELISA quantitative detection method and the sandwich ELISA quantitative detection kit of the GP73, the sensitivity and the specificity for detecting the GP73 are improved, high stability and reproducibility are guaranteed, the operating time is shortened and the cost is reduced.

Description

technical field [0001] The invention relates to an immunological detection method of serum markers for early diagnosis of liver cancer, in particular to a sandwich ELISA quantitative detection method of Golgi protein GP73 and a detection kit thereof. Background technique [0002] The liver is the largest organ in the human body, and liver disease is also the disease with the highest morbidity and mortality. Due to the strong compensatory ability of the liver, some injuries often have no clinical manifestations, resulting in a strong latent liver disease. When clinical manifestations appear, the liver disease is already at an advanced stage and it is difficult to effectively treat it. At present, there are many kinds of liver function tests that can be carried out clinically, but each test can only detect a certain function of a certain aspect of the liver. Up to now, there are still no indicators that can reflect most of the functions of the liver. Therefore, Often lead to ...

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
IPC IPC(8): G01N33/68
Inventor 彭涛古艳丽
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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