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

Anti-human ApoA1 monoclonal antibody for identifying HDL subclass in positive correlation with coronary heart disease and preparation and application thereof

A monoclonal antibody, cardiovascular and cerebrovascular disease technology, applied in the fields of application, botany equipment and methods, biochemical equipment and methods, etc., can solve the problem of inability to integrate, complicated and time-consuming operation, and failure to meet the needs of rapid and high-throughput clinical testing and other issues to achieve the effect of improving the consistency rate, high affinity and specificity

Pending Publication Date: 2021-08-17
DIASYS DIAGNOSTIC SYST SHANGHAI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current detection methods for subtypes of HDL, including ultracentrifugation, gel electrophoresis and nuclear magnetic resonance imaging (NMR), all require special and expensive equipment support, complex and time-consuming operations, and cannot be integrated into routine clinical detection systems. Rapid and high-throughput requirements for clinical testing

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
  • Anti-human ApoA1 monoclonal antibody for identifying HDL subclass in positive correlation with coronary heart disease and preparation and application thereof
  • Anti-human ApoA1 monoclonal antibody for identifying HDL subclass in positive correlation with coronary heart disease and preparation and application thereof
  • Anti-human ApoA1 monoclonal antibody for identifying HDL subclass in positive correlation with coronary heart disease and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation and purification of ApoA1 monoclonal antibody D11-6

[0043] 1. Materials: fusion protein 6×His-ApoA1, 8-week-old female BALB / c mice

[0044] 2. Methods and Results

[0045] 2.1 Antigen preparation

[0046] 2.1.1 Construction of human ApoA1 recombinant protein expression plasmid

[0047] The gene sequence (Gene ID: 335) of ApoA1 was obtained by querying in GenBank, and polymerase chain reaction (PCR) primers were designed according to the sequence. The DNA fragment of human ApoA1 gene was amplified using human cDNA as a template. The PCR product was detected by 1.0% agarose gel electrophoresis and the corresponding fragment was recovered with a gel recovery kit. The size of the ApoA1 gene fragment amplified by PCR was 875bp. The PCR product was ligated into the expression vector pCold TM II (the 6×His tag contained in the vector is used for purification), transformed into TaKaRaDH5α host bacteria, picked a single clone for plasmid extraction a...

Embodiment 2

[0065] Example 2: Determination of the variable region sequence of ApoA1 monoclonal antibody D11-6

[0066] 1. Materials: Trizol (Invitrogen), primers were synthesized by Sangon Bioengineering Company, reverse transcription and PCR reagents were purchased from TaKaRa Company.

[0067] 2. Methods and results:

[0068] 2.1 Total RNA extraction and cDNA first-strand synthesis

[0069] The hybridoma cells in the logarithmic growth phase were collected, and total RNA was extracted according to the operating procedure of the Trizol manual. The qualitative and quantitative identification of total RNA was performed using a spectrophotometer and agarose gel electrophoresis.

[0070] According to TaKaRa PrimeScript TM II 1st Strand cDNA Synthesis Kit Instructions Synthesize cDNA.

[0071] 2.2 Amplification and sequencing of gene fragments of ApoA1 monoclonal antibody D11-6 heavy chain variable region (VH) and light chain variable region (VL)

[0072] According to TaKaRa company's T...

Embodiment 3

[0074] Example 3: HDL subtype detection kit based on ApoA1 monoclonal antibody D11-6

[0075] 1. Materials: 96-well ELISA plate, rabbit anti-ApoA1 antibody, tris, PBS buffer (sodium chloride, potassium chloride, disodium hydrogen phosphate, potassium dihydrogen phosphate), bovine serum albumin (BSA ), ApoA monoclonal antibody D11-6, horseradish peroxidase (Horseradish Peroxidase, HRP) labeled goat anti-mouse secondary antibody, TMB chromogenic solution, reaction stop solution (2N sulfuric acid).

[0076] 2. Preparation and assignment of mixed serum calibrator: use serum samples from 20 healthy people, take an equal volume of serum for each case into a centrifuge tube (100 μL each), place at 4°C and mix thoroughly, then pack in 200 μL / tube, freeze Store in -80°C refrigerator. One was taken out the next day, and after thawing at 4°C, the ApoA1 immunoturbidimetric kit from Desai Company was used to assign values ​​to the mixed serum samples. The concentration of ApoA1 in the mi...

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
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The invention relates to an anti-human ApoA1 monoclonal antibody for identifying HDL subclasses positively correlated with coronary heart disease, and preparation and an application thereof. The invention discloses an ApoA1 monoclonal antibody D11-6, and the ApoA1 monoclonal antibody D11-6 can be used for specifically recognizing a specific antigen epitope of an ApoA1 protein. The ApoA1 monoclonal antibody D11-6 is secreted by a hybridoma cell strain D11-6, the hybridoma cell strain D11-6 is preserved in the China Center for Type Culture Collection (CCTCC), the preservation date is April 26, 2021, the preservation number is CCTCC NO: C202187, and the preservation address is Wuhan University, China. The ApoA1 monoclonal antibody D11-6 disclosed by the invention is used for developing a kit suitable for clinical high-flux immunoturbidimetry automatic detection, so that risk prediction and clinical monitoring of cardiovascular and cerebrovascular diseases can be realized, and a foundation is further laid for clarification of occurrence and development mechanisms of the diseases.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to an anti-human ApoA1 monoclonal for identifying HDL subtypes positively related to coronary heart disease and its preparation and application, in particular to an anti-human apolipoprotein A1 (ApoA1) mouse-derived monoclonal antibody D11 -6 and the hybridoma cell line secreting the monoclonal antibody, and the application of the monoclonal antibody in clinically detecting the content of ApoA1 protein in human peripheral blood to predict the risk of cardiovascular and cerebrovascular diseases. Background technique [0002] Due to the long course of the disease, the high cost of treatment, and the high rate of death and disability, coronary heart disease brings great life threats and economic burdens to patients; it occupies a large amount of social medical resources and becomes a hidden danger to social development. Therefore, early detection and diagnosis are of great significanc...

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): C07K16/28C12N5/20C12N15/13G01N33/68G01N33/577C12R1/91
CPCC07K16/2878G01N33/6893G01N33/577C07K2317/33C07K2317/34C07K2317/56C07K2317/92G01N2333/775G01N2800/324G01N2800/2871
Inventor 张杰吕良杰张艳邹艳芳
Owner DIASYS DIAGNOSTIC SYST SHANGHAI
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